Domain structure of mitochondrial and chloroplast targeting peptides
|
|
- Åke Adam Eriksson
- för 5 år sedan
- Visningar:
Transkript
1 Eur. J. Biochem. 180, FEBS 1989 Domain structure of mitochondrial and chloroplast targeting peptides Gunnar von HEIJNE, Johannes STEPPUHN and Reinhold G. HERRMANN Department of Molecular Biology, Karolinska Institutet, Center for Biotechnology, Huddinge Hospital Botanisches Institut der Ludwig-Maximilian-Universitat, Munchen (Reccived October 18/November 28, 1988) - EJB Representative samples of mitochondrial and chloroplast targeting peptides have been analyzed in terms of amino acid composition, positional amino acid preferences and amphiphilic character. No highly conserved homology blocks are found in either class of topogenic sequence. Mitochondrial-matrix-targeting peptides are composed of two domains with different amphiphilic properties. Arginine is frequently found either at position - 10 or - 2 relative to the cleavage site, suggesting that some targeting peptides may be cleaved twice in succession by two different matrix proteases. In stroma-targeting chloroplast transit peptides three distinct regions are evident: an uncharged amino-terminal domain, a central domain lacking acidic residues and a carboxy-terminal domain with the potential to form an amphiphilic P-strand. Targeting peptides that route proteins to the mitochondrial intermembrane space or the lumen of chloroplast thylakoids have a mosaic design with an aminoterminal matrix- or stroma-targeting part attached to a carboxy-terminal extension that shares many characteristics with secretory signal peptides. Both mitochondria and chloroplasts import most of their proteins from the cytoplasm in the form of preproteins with amino-terminal extensions (targeting or transit peptides). In most cases, the targeting peptides are removed by intraorganellar proteases during of shortly after import. Since both mitochondrial and chloroplast targeting peptides can direct the import of normally cytoplasmic proteins they should carry most of the information required for correct sorting; yet, the only common themes suggested so far are the possible existence of an amphiphilic a-helix in mitochondrial targeting peptides [l - 31 and three homology blocks of conserved amino acids in chloroplast transit peptides [4-51. In this paper, we present the results of an extensive analysis of collections of nonhomologous mitochondrial targeting (mtp) and chloroplast targeting (ctp) peptides. We find no signs of conserved homology blocks in either class of sequence. mtps are composed of two structurally distinct domains : an amino-terminal domain with amphiphilic a-helical character and a carboxy-terminal domain with different amphiphilic properties. In the latter domain, Arg residues are frequently found either around position -2 or position - 10, counting from the cleavage site. ctps also show signs of a domain structure with an uncharged amino-terminal part, a central nonamphiphilic part lacking acidic residues and a carboxy-terminal part that is predicted to form an amphiphilic 6-strand. We suggest that ctps, unlike mtps, will not spontaneously integrate into lipid bilayers to any significant degree; rather, we favour a model where they bind either to the membrane surface and/or to specific receptors on the chloroplast. Correqmndence to G. von Heijne, Institutionen for Moleky- Iiirbiologi, Huddinge sjukhus, K87, S Huddinge, Sweden Abbreviations. mtp, mitochondrial targeting peptide; ctp, chloroplast transit peptide. MATERIALS AND METHODS Sequence samples A total of 37 mitochondrial targeting peptides (mtps) with known cleavage sites (matrix protease) were collected from the literature (Table 1). This collection was further subdivided into a sample containing sequences from Saccharomyces and Neurospora, and a sample containing sequences from higher eukaryotes. The two samples were analyzed separately as well as pooled. Likewise, a total of 18 chloroplast transit peptides (ctps) with known cleavage sites (stromal protease) were analyzed (Table 1). For the analysis of the amino-terminal region, an additional eight ctps with unknown cleavage sites were included in the sample (Table 1). Control samples of mature, imported mitochondrial or chloroplast proteins included all soluble or peripheral membrane proteins of known sequence (with their targeting sequences removed) from Table 1. For comparison with the amino-terminal region of imported mitochondrial proteins, residues 1-15 were removed from all sequences in the control sample. Apolar regions from secretory signal peptides were obtained from the SIGPEP database [36]. Residues -13 to -7 from a total of 94 prokaryotic signal peptides and residues - 13 to - 6 from a total of 472 eukaryotic signal peptides, all with known cleavage sites, were pooled. Hydrophobic moment analysis Hydrophobic moment analysis was performed according to [37]. The window size was 11 residues for mtps and 10 residues for ctps. The statistical significance of peaks in the hydrophobic moment vs. 6 plots was assessed by comparison with the hydrophobic-moment profiles for control samples of
2 cn w (T\ Table 1. Mitochondriul-targeting peptides and chloroplast transit peptides analyzed in this paper Known cleavage sites are indicated by... PS. photosystem; RuBisCO, ribulosc bisphosphate carboxylaseioxygenase; EPSP, 5-enolpyruvylshikimate-3-phosphate synthase Source Protein Sequence Reference Mitochondrial-targeting peptides Sacchuromyces ATPase F, p-subunit ATPase Fo subunit 4 Cytochrome oxidase IV Cytochrome oxidase V Cytochrome oxidase VI Cytochrome oxidase VIII Superoxide dismutase (Mn) Stabilizing factor 9-kDa protein Rieske Fe-S protein C1-tetrahydrofolate synthase Dihydrolipoamide dehydrogenase Neurospora Bovine Chicken Human Maize Rat Cytochrome oxidase IV Cytochrome oxidase V Proteolipid Adrenodoxin Adrenodoxin reductase ATPase inhibitor Cytochrome oxidase IV Cytochrome P-450 (side-chain cleavage) Cytochrome P-450 (1 1 B) ATP synthase F6 Oligomycin sensitivity conferral protein Proteolipid PI 5-Aminoleavulinate syiithase Aspartate aminotransferase ATPase F1 B subunit Lipoamide dehydrogenase Superoxide dismutase (Mn) Ornithine transcarbamylase Pyruvate dehydrogenase a-subunit Pyruvate dehydrogenase /?-subunit Superoxide dismutase (Mn) Carbamyl phosphate synthetase I Malate dehydrogenase Serine: pyruvate arninotransferase Succinyl-CoA syntbetase, mubunit Aldehydedehydrogenase MVLPRLYTATSRAAFKAAK^QSAPLLSTSW MSMSMGVRGLALRSVSKTLFSQGVRCPSMVIGARY-MSSTPEKQTD MLSLRQSIRFFKPATRTLCSSRYLL-QQKPVVKTAQ MLRNTFTRAGGLSRITSVRF^AQTHALSNAA MLSRAIFRNPVINRTLLRARPGAYHATRLTKNTFIQSRKY-SDAHDEETFE MLCQQMIRTTAKRSSNIMTRPllMKRS^VHFKDGVYEN MFAKTAAANLTKKGGLSLLSTTARRT-KVTLPDLKWD MLNRCISRNTRLPVNLRIASRFY-SDGPLGGAGP MLGIRSSVKTCFKPMSLTSKRLISQSLLAS-KSTYRTPNFD MLSRLSLLSNSRAFQQARWRIYRLKVSPTVHASQ-YHILSGRKLA MLRIRSLLNNKRAFSSTVRTL-TINKSHDVVI... RAPALRRSIATTVVRC-NAETKPVPPH MLRTPTVSALVRNVAVRAAKPTMAVRA^ASTMPISNPT MASTRVLASRLASQMAASAKVARPAVRVAQVSKRTTIQTGSPLQTLKRTQMTSIVNATTRQAFQKRA~YSSEIAQAMV MAARLLRVASAALGDTAGWRLLVRPRAGAGGLRGSRGPGLGGGAVATRTLSVSGRAQ~SSSEDKITVH MAPRCWRWWPWSSWTRTRLPPSRSIQNFGQHF~STQEQTPQIC MAATALAARTRQAVWSVWAMQGRGF*GSESGDNVRS MLATRVFSLIGRRAISTSVCVR-AHGSVVKSED MLARGLPLRSALVKACPPILSTVGEGWGHHRVGTGEGAG^ISTKTPRPYS MALWAKARVRMAGPWLSLHEARLL^GTRGAAAAPKA MILQRLFRLSSAVQSAISVSWRRNIGITAVAF^NKELDPVQKL MAALAVSGLSQQVRCFSTSVVRP^FAKLVRPPVQ MQTTGALLISPALIRSCTRGLIRPVSASFLSRPEIQSVQPSYSSGPLQVARREFQTSVVSR~DIDTAAKFIG MEAVVRRCPFLARVSQAFLQKAGPSLLFYAQHCPKMMEAAGQ-QVEETPAAQP MALLQSRLLLSAPRRAAATARA^SSWWSHVEMG MTSLWGKGTGCKLFKFRVAAAPASGALRRLTPSASLPPAQLLLRAVRRRSHPVDYAAQ~TSPSPKAGAA MQSWSRVYCSLAKRGHFNRISHGLQGLSAVPLRTY^ADQPIDADVT MLSRAVCGTSRQLAPAIJGYLGSRQ~KHSLPDLPYD MLFNLRILLNNAAFRNGHNFMVRNFRCGQPLQ*NKVQLKGRDL MRKMLAAVSRVLSGASQKPASRLLVASRN-FANDATFEIK MAAVSGLVAETPSEVSGLLKRRFHWTAPAAAVQVTVRDAIN MALRTLASKKVLSFPFGGAGRPLAAAASARG^VTTVTTVTLPDLS MTRILTACKVVKTLKSGFGLANVTSKRQWDFSRPGIRL^LSVKAQTAHI MLSALARPVGAALRRSFSTSAQ"^AKVAVLGASG MFRMLAKASVTLGSRAASWVRNMG^SHQLLVPPPE MVSGSSGLAAARLLSRTFLLQQNGIRH-GSYTASRKNI MLRAALSTARRGPRLSRLL-SAAATSAVPA
3 [771 ~781 [821 ~ 3 1 ~ 5 1 Spinach Chloroplast transit peptides 10-kDa protein ATPase &subunit Ferrodoxin oxidoreductase Light-harvesting-complex protein I1 MATSVMSSLSLKPSSFGVDTKSAVKGLPSLSRSSASFTVRA^SGVKKIKVDK MAALQNPVALQSRTTTAVAALSTSSTTSPPKFSLSFSSSTATFNPLRLKILTASKLTAKPRGGALGTRM-VDSTASRYAS MTTAVTAAVSFPSTKTTSLSARSSSVISPDKISYKKVPLYYRNVSATGKMGPIRA-QIASDVEAPP MASSTMALSSPSLAGKAVKLGPTASEIIGEGRIT^MRKTAGKPKT 1691 [331 [701 Wedel, personal Rieske Fe-S protein Small subunit of RuBisCO RuBisCO activase Nitrite reductase Acyl carrier protein I PS 1-3 PS 1-5 PS 1-6 Glutamine synthase 2 Thioredoxin F PS I1 20-kDa protein communication MIISIFNQLHLTENSSLMASFTLSSATPSQLCSSKNGMFAPSLALAKAGRVNVLISKERIRGMKLTCQ~ATSIPADNVP [711 MASSVLSSAAVATVSRTPAQASM~APFTGLKSTVGFPATKNDDITSLASNGGRVQC-MKVWPTQ~MK [721 MATAVSTVGAATRAPLNLNGSSAGASVPTSGFLGSSLKKHTNVRFPSSSRTTSMTVKA~AENEEKNTDK [731 MASLPVNKIIPSSTTLLSSSNNNRRNNSSIR-CQKAVSPAAE [741 MASLSATTTVRVQPSSSSLHKLSQGNGRCSSIVCLDWGKSSFPTLRTSRRSFISA-AKKETIDKVC [751 MASIASSVAVRLGLTQVLPNKNFSSPRSTRLVVR^AAAPA unpublished results MAAATASLSSTLLAPCSSKQPQPQQQHQHQQLKCKSFSGLRPLKL~ISSNNSSSSLSMSSARRSMTCRA-ELSPSLVISL unpublished ~761 MASLATLAAVQPTTLKGLAGSSIAGTKFTSARRQSFKL~VRSGAIVA-KYGDKSVYFD results MAQILAPSMQCQMKLSKGLTSSMTPSPWTSILLKQGQKGSIKCSTKFRVCAS~QSDHG unpublished results MALHLSLSHQSWTTPAHPITSSDPTRSSVPGTGLSRRVDFLGSCKINGVFVVKRKDRRRMRGGE unpublished results MAAATSATAIVNGFTSPFLSGGKKSSQSLLFVNSKVGAGVSTTSRKLVVVAAAAAPKKSWIPAVKGGGNFL unpublished results Silene Ferredoxin Pea Early light-induced protein Heat-shock protein Gin synthase Tomato PS 1-2 PS I1 Chlorophyll ah apoprotein Light-harvesting-complex protein I Brassica Acyl carrier protein Chlamydomonas Small subunit of RuBisCO Arabidopsis EPSP Petunia EPSP MASTLSTLSVSASLLPKQQPMVASSLPTNMGQALFGLKAGSRGRVTAM~ATYKVTLITK MAVSSCQSIMSNSMTNISSRSRVNQFTNIPSVYIPTLRRNVSLKVRSMAEGEPKEQS MAQSVSLSTIASPILSQKPGSSVKSTPPCMASPLRRQLPRLGLRNVRAQAGGDGDN MSSLSDLINFNLSDSTEKIIAEYIWVGGSGIDIRSKARTLPGPVSDPAKLPKWNY MAMATQASLFTPPLSVPKSTTAPWKQSLVSFSTPKQLKSTVSVTRPIRAM-AEEAPAATEE MATSMALSSSTFAGKAVKLSPSSSEITGNGRVTMRKTATKAKPASSGSPWYGPDRVKY MASNTLMSCGIPAVCPSFLSSTKLSKFAAAMPVSVGATNSMSRFSM MSTTFCSSVSMQATSLAATTRISFQ~PALVSTTNLSFNLRRSIPTRFSISC-AAKP~TVEKV ~ 4 1 MAVIAKSSVAAVARPARSSVRPMAALKPAVKAAPVAAPAEAND-MMVWTPVNNK [791 [801 [811 [
4 randomly scrambled copies of each wild-type 11- or 10- residue sequence using a one-sided t-test. An amphiphilic a-helix is characterized by a strong hydrophobic-moment component for an angle 6 = " between successive residues and an amphiphilic B-sheet has a corresponding strong component for 6 = " [7]. Amino-acid-frequency analysis Overall amino acid distributions in the various samples were compared using a 1' test (19 degrees of freedom). When two distributions were found to be significantly different (P < by this method, the counts for individual amino acids were compared again using a 1' test (one degree of freedom). The statistical significance of peaks in the position-specific distribution of individual amino acids was assessed by calculating, from a binominal distribution, the probability that an equal or greater number of the residue in question would be found when n residues (n = number of sequences in the sample) were drawn at random from a pool of amino acids with average frequencies the same as those found in the full mtp or ctp samples. RESULTS Mitochondria1 targeting peptides In a previous study, we showed that mtps are enriched for Arg, Leu and Ser, contain very few Asp and Glu and have the potential to fold into amphiphilic a-helices [2]. We have now extended this study to include a total of 37 mtps with known cleavage sites (Table 1). mtps from lower eukaryotes (Saccharomyces and Neurospora, 14 entries) have been analyzed separately from mtps from higher eukaryotes (mainly mammals, 23 entries). Overall amino acid composition qf mtps With the larger and more well-defined collection now available, we have re-evaluated the amino acid preferences found in our earlier study. As shown in Table 2, mtps contain significant more Ala, Leu, Arg and Ser and less Asp, Glu, Ile and Lys residues than the control sample of imported, soluble mitochondria1 proteins (P < in all cases). The reduction in Ile and Lys is only apparent in mtps from higher eukaryotes. mtps from lower eukaryotes are also significantly enriched for Thr (data not shown). The C-terminal residues have difierent umphiphilic properties than the N-terminal domain Since positively charged, amphiphilic a-helices have been suggested to be important for the function of mtps, we first wanted to know whether these amphiphilic sections were distributed throughout the mtps, or confined to a smaller part. The mtps in our sample were aligned with coincident cleavage sites, and the mean a-helical hydrophobic moment (6 = 95") for successive 11-residue segments was determined (Fig. 1 A). As a control, the amino acids in each 11-residue segment in each sequence were scrambled and reassembled into 100 random segments and the same analysis was performed on this randomized sample. From the figure it is clear that the carboxy-terminal residues do not consistently fold into cr-helices with a higher hydrophobic moment than the Table 2. Amino ucid,frequencies Statistically significanl differences (P < text for description of peptidcs analyzed Amino Mitochondrial Chloroplast pcptides acid peptidcs A C D E F G H I K L M N P Q R S T V W Y Total are indicated by *. See mtp mature ctp -lo/-1 N 3-10 mature 0.141* * * * * * * * * * * * 0.192* * * * * * * controls. This was confirmed using shorter windows of 8-10 residues (not shown). Residues -24 to -15, on the other hand, have a high mean a-helical hydrophobic moment. In order to characterize the carboxy-terminal region further, we calculated the mean hydrophobic moment of the samples as a function of 6 for residues - 1 I to - 1 (Fig. 1 B). A significant peak was apparent around 6 = 75 ' (P < lo-'). Also shown in Fig. 1 B is the hydrophobic-moment profile for residues -18 to -8, with a typical amphiphilic a-helix peak at 6 = 95" (P < lop3). The transition between the two regions is readily apparent in the 6 = 75" and 6 = 95" profiles in Fig. 1 A. Similar results were obtained when the mtps from lower and higher eukaryotes were analyzed separately (not shown). These observations suggest that the carboxy-terminal residues in mtps do not, as a rule, tend to form classical amphiphilic a-helices. The possible significance of the 6 = 75" structure is discussed further below. Arg is enriched around positions - 2 and - 10 Since the cleavage of matrix-targeting mtps from mature proteins is highly specific, it seemed likely that distinct patterns of individual amino acids might be found in the region close to the cleavage site. We thus calculated the incidence of each of the 20 amino acids as a function of position relative to the cleavage site (located between positions - 1 and + 1). Since the data base is still fairly small, only very strong position-specific preferences can be found at this stage. Nevertheless, a significant enrichment of Arg residues is found at position -2 (P < lop3) and at position -10 (P < Fig.2). Less significant peaks are found in the neighboring positions, - 3 and - 11 (P < 0.05). The - 2 peak is apparent
5 539 Fig. 1. Mean normalized hydrophobic moment of mtps as a function of 6 and of position relative to the cleavage site. (A) Mean normalized hydrophobic moment of mtps as a function of position relative to the cleavage site (located between positions -1 and +I). The mean hydrophobic moment (p) for an 1 I-residue window starting at the indicated position was normalized by first subtracting the mean value (p)5cr calculatcd for a control sample consisting of 100 randomly scrambled copies of each of the wild-type 11-residuc stretches contributing to a given point and then dividing by the standard deviation u of the calculated (p) value. The shaded area indicates 2 standard deviations in the determination of (p);(0) 6 = 95"; (+) 6 = 75". (B) Mean normalized hydrophobic moment of mtps as a function of 6 for segments - 11 to - 1 (0) and - 18 to -8 (W). Normalization was carried out as in A. The shaded area indicates f 2 standard deviations in the determination of (p) Table 1 were compared with the composition of a sample including all remaining residues, i.e. from residue 16 to the carboxy terminus (excluding the integral membrane proteins, see Materials and Methods). Two differences were observed : Pro and Ser are significantly more abundant in the aminoterminal region of the mature proteins (So/, vs 5% Pro and 9% vs 6% Ser, P < for both). In summary, at least two domains can be identified in mtps. The adjoining amino-terminal region of the mature proteins also seems to have a distinct amino acid composition I I6 +21 Position Fig. 2. Positional distrihufion of Arg residues in nztps. The cleavage site is between positions - 1 and + 1 (arrow) Chloroplasl targeting peptides A total of 18 stroma-targeting ctps with known cleavage sites and an additional eight sequences where the stromal cleavage sites have not been determined (Table 1) were analyzed using the same methods as above. in mtps from both lower and higher eukaryotes, whereas the - 10 peak is much more prominent in the sample from higher eukaryotes (not shown). Interestingly, mtps with Arg at position - 2 do not seem to be enriched for Arg at position - 10 and vice versa (data not shown). Furthermore, out of 16 sequences with Arg in position - 10, all but one have an apolar residue in position -8 (mostly Phe, Ile or Leu). Arginine residues elsewhere in the rntps do not show this bias towards apolar residues in their downstream next-nearest-neighbor position and mtps that do not have Arg in position -10 are not enriched for apolar residues in position - 8. We also compared the overall amino acid composition of the amphiphilic amino-terminal regions (up to and including residue - 17) and the carboxy-terminal regions (residues - 13 to - 1). No significant differences were observed. The,first residues of' mature, imported mitochondria1 proteins are rich in Pro and Ser The amino acid composition of the 15 amino-terminal residues of the mature *mitochondria1 proteins shown in CTPS contain.few acidic residues, but many Ser and A h The overall amino acid composition of the ctps were compared with the composition of mature chloroplast proteins (Table 2). Significant differences (P < lop3) were found for Ser and Thr, which are more abundant in the ctps (19% vs 6% for Ser, 9% vs 5% for Thr) and for Asp, Glu and Tyr which are almost completely absent from the ctps. In contrast to the mtps, however, Arg and Leu were not found to be enriched in the chloroplast transit peptides. It has been suggested that ctps contain three fairly wellconserved 'homology blocks' [4]: an amino-terminal block with the consensus sequence MAXSXMXSS (where X denotes the possible presence of an undefined residue), a central block PXFXGXK, and a carboxy-terminal block GXGRV just before the cleavage site. These blocks were found by analyzing six homologous ctps from light-harvesting complex protein I1 polypeptides, six homologous ctps from ribulose-bisphosphate carboxylase/oxygenase small subunit polypeptides, one ferredoxin, and one plastocyanin ctp. We have searched our collection of nonhomologous ctps and found no significant conservation of any of these blocks except for the-amino-terminal MA dipeptide. Neither have we
6 540 Fig. 3. Mean normulized hydrophobic moment of chlorophst transit peptides. (A) Shows the mean normalized hydrophobic moment as a I'unction of 6 for the rcgion - 10 to -1, and (B) is the positional plot for 6 = 165" (cf. Fig.1) found any evidence that a critical serine at position - 8 relative to the cleavage site [6] is conserved. We conclude that ctps do not contain regions of highly conserved amino acids. The amino-terminal 10 residues,form a distinct domain The amino-terminal region of the ctps in our sample is significantly different (P < from the remainder of these sequences in overall amino acid composition: there are very few charged amino acids (Arg, Lys, Asp, Glu) and very few Pro and Gly among the first 10 residues (Table 2). In addition, the residue next to the initiator Met is almost invariably an alanine (22 cases out of 26). Serine, which is present at a level of around 20% in thc ctps, is not specifically enriched in any one position in the amino-terminal region. Hydrophobicmoment analysis does not reveal any amphiphilic structures when applied to the sample of N-terminal-aligned ctps. The region next to the cleavage site is a potential amphiphilic P-strand Hydrophobic moment analysis reveals that the last 10 residues of stroma-targeting ctps have a very pronounced peak (P < lo-') in the hydrophobic moment plot for an angle 6 = 165" between successive residues (Fig.3A). This value of 6 corresponds to the slightly curved amphiphilic P-strand normally found in P-sheets [7]. As shown in Fig. 3 B, the 6 = 165" structure is only found in the carboxy-tcrminal region. No other amphiphilic structure is apparent in the C-termiaalaligned sample (data not shown). It is interesting to note that thylakoid-targeting ctps (see below) often seem to have an intermediate cleavage site close to a similar stretch of amphiphilic P-strand [8]. It is thus possible that the same stromal protease cleaves both kinds of ctps. The amino acid composition of the carboxy-terminal 10 residues is also different from the more amino-terminal parts. Specifically, Arg is enriched in this region (14% vs 3%, P < and Leu is found only infrequently (2% vs lo%, P < Table 2). Although the ctp sample is rather too small for the reliable identification of position-specific amino acid preferences, it is apparent that turn-inducing residues (Gly, Ser, Asp, Asn, Pro) [9] are largely absent from positions - 3 to + 1, i.e. from the immediate vicinity of the stromal cleavage site. Ala, on the other hand, occurs rather frequently in this region (data not shown). Targeting to the mitochondria1 intermembrane space and to the thylakoid lumen depends on mosaic targeting peptides with signal-peptide-like extensions Both the mtps and ctps analyzed above target proteins for import through two membranes into the matrix of mitochondria or the stroma of chloroplasts. However, some proteins are routed to other suborganellar locations such as the mitochondrial intermembrane space or the chloroplast thylakoid-membrane system. It has been proposed that intermembrane-space proteins are first imported into the matrix compartment and then re-exported through the inner membrane [lo]. A similar model may be applicable to proteins of the thylakoid lumen and to some thylakoid-membrane proteins: they are first imported into the stromal compartment and then further routed into the thylakoid [I 11. A collection of targeting signals from such proteins is shown in Fig. 4. It is immediately obvious that these signals are mosaic structures with a typical amino-terminal matrix-targeting intp or stroma-targeting ctp attached to a carboxy-terminal apolar domain. Indeed, this latter region is very similar to known secretory signal peptides involved in routing proteins into the secretory pathway of both prokaryotic and eukaryotic cells [12]. Signal peptides are characterized by a short aminoterminal positively charged region, a central apolar region some 7-15 residues long, and a carboxy-terminal region typically 5 - h residues long and with small, uncharged amino acids in positions -3 and -1, counting from the cleavage site. Assuming that the mosaic targeting peptides are first cleaved just before the apolar stretch upon import into the matrix or stroma [8], bona fide amino-terminal signal peptides would be generated. In the case of the thylakoid-targeting peptides this analogy extends all the way up to and including the final cleavage site (A-X-AJ), whereas the mitochondrialintermembrane-space-targeting peptides seem to be cleaved at a final site which does not always follow the (-3, - 1) pattern (Fig. 4). From preliminary data (obtained in collaboration with Dr C. Robinson) it appears that at least some thylakoid-targeting ctps are first cleaved in, or close to, a potential amphiphilic P-strand similar to that found in stroma-targeting ctps, thus exposing the signal-peptide-like structure. It is interesting to note that the apolar region of the intraorganellar signals differs significantly in amino acid composition from the corresponding regions in both prokaryotic and eukaryotic signal peptides. The typical apolar region of
7 Yeast cytochrome c1 [87] MFSNLSKRWAQRTLSKSFYSTATGAASKSGKLTQKLVTAGVAAAGITASTLLYADSLTAEA#MTAAEHGLHA Yeast cytochrome hz [88] MLKYKPLLKISKNCEAAILRASKTRLNTIRAYGSTVPKSKSFEQDSRKRTQSWTALRVGAILAATSSVAYLNWHNGQIDN#EPKLDMNKQKN Yeast cytochrome c peroxidase [89] MTTAVRLLPSLGRTAHKRSLYLFSAAAAAAAAAATFAYSQSHKRSSSSPGGGSNHGWNNWGKAAALAS#TTPLVHVASV Human cytochrome c1 [90] MAAAAASLRGVVLGPRGAGLPGARARGLLCSARPGQLPLRTPQAVALSSKSGLSRGRKVMLSALGMLAAGGAGLAVALHSAVSA#SDLEVHPPSY Spinach 16-kDa protein [91] MAQAMASMAGLRGASQAVLEGSLQISGSNRLSGPTTSRVAVPKMGLNIRAQQVSAEAETSRRAMLGFVAAGLASGSFVKAVLA#EARPIVVGPP Spinach 23-kDa protein [91] MASTACFLHHHAAISSPAAGRGSAAQRYQAVSIKPNQIVCKAQKQDDNEANVLNSGVSRRLALTVLIGAAAVGSKVSPADA#AYGEAANVFG Spinach 33-kDa protein [8] MAASLQASTTFLQPTKVASRNTLQLRSTQNVCKAFGVESASSGGRLSLSLQSDLKELANKCVDATKLAGLALATSALIASGANA#EGGKRLTYDE Spinach plastocyanin [92] MATVASSAAVAVPSFTGLKASGSIKPTTAKIIPTTTAVPRLSVKASLKNVGAAVVATAAAGLLAGNAMA#VEVLLGGGDGSLAFLPGDFS Spinach CFo-2 [Steppuhn et al., unpublished results] MANMLVASSSKTLPTTTTTTITPKPKFPLLKTPLLKLSPPQLPPLKHLNLSVLKSAAITATPLTLSFLLPYPSLA#EEIEKASL~D Spinach photosystem 1-4 protein [76] MSFTIPTNLYKPLATKPKHLSSSSFAPRSKIVCQQENDQQQPKKLELAKVGANAAAALALSSVLLSSWSVAPDAAMA#DIAGLTPCKE Chlanzydomonas reinhardtii oxygen-evolving complex 2 (33 kda) [93] MATALCNKAFAAAPVARPASRRSAVVVRASGSDVSRRAALRGFAGAAALVSSSPANA#AYGDSANVFG Chlamydomonas reinhardtii Cu(1I)-repressible cytochrome c [21] MLQLANRSVRAKAARASQSARSVSCAAAKRGADVAPLTSALAVTASILLTTGAASASA#ADLALGAQVF Pea cytochrome f [94] MQLTRNAFSWIKKEITRSISVLLMIYIITRAPISNA#YPIFAQQGYE Fig.4. A collection ofintru-organellar sorting signals. The location of the photosystem 1-4 protein is not known, but the structure of its transit peptide suggests that it is lumenal. Final cleavage sites are indicated by #. References are given in square brackets, apolar regions are underlined ' w 0 g c A C D E F G H I K L M N P O R S T V W Y Residue Fig. 5. Amino acid content of the apolur regions underlined in Fig.4. Intra-organellar sorting signals (black bars); bacterial secretory signal peptides (white bars) and eukaryotic secretory signal peptides (grey bars) a eukaryotic signal peptide has some 40% Leu, 10% Ala and 40% other apolar residues; that of a prokaryotic signal peptide has 25% Leu, 20% Ala and 40% other apolar residues; and the apolar regions of the intra-organellar signals have 20% Leu, 35% Ala and 20% other apolar residues (Fig. 5). DISCUSSION Mitochondria1 and chloroplast targeting peptides contrasted As shown above, neither mtps nor ctps contain stretches of highly conserved sequence; rather, they are remarkably variable in their amino acid sequence. Nevertheless, they seem to be built from two (mtp) or three (ctp) structurally distinct domains and they have overall amino acid compositions that are very different from average soluble proteins. In terms of gross amino acid content, both mtps and c' 1 1's lack acidic residues, but beyond this they are different: mtps are mainly enriched in Arg (14%), Ser (11Y0) and Ala (14%), whereas ctps are characterized by a high percentage of Thr (9%) and an extremely high content of Ser (19%). In fact, it is possible to discriminate efficiently between mtps and ctps by simply checking whether the fraction of Ser in the targeting peptide,f,,,, is smaller or greater than xfarg. More than 90% of the mtps can be told apart from more than 90% of the ctps in this way. In mtps, two domains can be discerned: an amino-termind region, that can potentially fold into an amphiphilic a- helix (6 = 95"), and a carboxy-terminal domain of residues that is characterized by a strong component of the hydrophobic moment for 6 = 75". Within this latter domain, Arg is very often found at positions -2 and -10. The 6 = 75" structure could correspond to an a-helix with a clockwise spiralling ridge of apolar (or polar) amino acids
8 542 A B Fig.6. Helical net plots of residues - 18 to - 1 of the mtps from the Saccharomyces Rieske Fe-S protein (A) and Neurospora rytochrornc. oxidase IV (B). Putative apolar i, i + 5 ridges are shaded and positively charged residues are highlighted. The cleavage site is at the top of the diagrams. Note that two identical copies of the helical net have been placed side by side to allow the apolar i, i + 5 spirals to be seen more clearly involving residues i, i + 5, i + 10, etc. In fact, such a ridge can be seen on helical net plots of the carboxy-terminal region in many mtps (Fig. 6) and could be an indication that helix/ helix packing [I31 might take place between mtps and the matrix protease responsible for cleavage. It is also noteworthy that the amino-terminal regions of mature imported mitochondrial proteins are rich in Pro and Ser. Conceivably, this could be related to the cleavage specificity (see below). Stroma-targeting ctps seem to have at least three distinct regions. At their amino-terminal they have a stretch of some 10 amino acids that is mostly uncharged and also contains few Pro and Gly residues (there is no corresponding domain in mtps). The residue next to the initiator Met is almost invariably Ala (in mtps this residue is often Leu, 40% of all sequences, but almost never Arg, 3% of all sequences). This probably means that the initiator Met is normally removed in ctps but not in mtps [14, 151. The central region is of variable length, is rich in Ser, but contains few if any acidic residues. Finally the carboxy-terminal region (8-10 residues), contains an increased proportion of Arg residues and can be folded into an amphilic /?-strand. The number of nonhomologous sequences with known cleavage sites is not yet sufficiently large to draw any strong conclusions regarding positional amino acid preferences in the vicinity of the cleavage site, but turn-promoting residues seem to be avoided at positions - 3 to + 1. We also note that the amino- and carboxy-terminal regions of ctps seem to be more sensitive to deletions than the central region [6, 161. The CdrbOXy-terminal amphiphilic /?-strand would seem likely to be involved in interactions with the stromal protease rather than with the chloroplast envelope and, except for the uncharged but not very hydrophobic amino-terminal domain, there are no other indications from the sequence data that ctps can form structures that might partition into lipid bilayers. mtps, on the other hand, due to their amphiphilic character, interact strongly with lipid monolayers and bilayers in model systems [l, 171. Possibly, the net positive charge of ctps could induce a loose association with the surface of a negatively charged membrane, but in addition some kind of receptor-mediated mechanism must be imagined. The absence of any significant overall enrichment in positively charged residues (in contrast to the high incidence of Arg in mtps) may be related to the fact that, unlike mitochondria, protein import into chloroplasts does not require an electrochemical gradient across the inner envelope membrane [I 81. Another possibility is that Arg residues in mtps may bind specifically to cardiolipin present in the mitochondrial but not the chloroplast envelope [19, 201. A remarkable exception to these clear-cut differences between mtps and ctps is provided by the Chlamydomonas Cu(I1)-repressible cytochrome c ctp (Fig.4). This is a mosaic sorting signal targeting the protein to the thylakoid lumen [21]. However, the amino-terminal, presumably stroma-targeting, part would be classified with the matrix-targeting mtps by all criteria discussed above: high Arg and modest Ser content, Met-Leu amino-terminal sequence and a high hydrophobic moment for 6 = 95" but not for 6 = 165" (data not shown). It should be interesting to study the behavior of this protein in in vitro import assays. Cleavage sites in mtps and ctps Our results show that the specifications of the cleavage sites in mtps and ctps do not reside in highly conserved consensus patterns of amino acids. The amphiphilic b = 75" structure found just before the cleavage site in mtps and the 6 = 165" structure found in a similar position in ctps most probably contribute to cleavage-site recognition. The high content of Pro and Ser in the region immediately downstream of the mtp cleavage site suggests that the possibly a-helical mtp is normally followed by a rather loosely structured piece of chain; the transition point could be a recognition element for a matrix protease. Arg residues around positions -2 and - 10 also seem to play an important role in defining the precise cleavage site in mtps (the prevalence of Arg in position -2 has been noted before [22]). It has recently been shown that human mitochondria contain two distinct matrix-localized processing activities, proteases I and I1 [23]. Protease I seems to cleave all matrix-targeting mtps, whereas protease 11 cleaves only some mtps at a second site, subsequent to cleavage by protease 1. Protease I thus generates either the mature protein or an intermediate that is further processed by protease 11. In the
9 543 two cases where a protease I intermediate has been documented (rat ornithine transcarbamylase and malate dehydrogenase), the first cleavage takes place between residues -9 and - 8 (Arg-AsnJPhe) for ornithine transcarbamylase or between - 10 and -9 (Arg-ArgJSer-Phe) for malate dehydrogenase [24, 251. In both cases, residue -10 is an Arg and residue -8 is an apolar Phe. Neither sequence has Arg in position -2 relative to the amino terminus of the mature protein (Val-GlnJSer and Asn-AsnlAla). This fits quite well with the findings reported here: almost all mtps have either an Arg in position -2 or an Arg in position -10 followed by an apolar residue in position -8. Few have Arg in both positions - 10 and -2, and those that lack an Arg in position - 10 also tend to lack an apolar residue in position - 8. Possibly, most or all mtps with an Arg in position -10 followed by an apolar residue in position - 8 and lacking an Arg in position - 2 are cleaved in two steps: first, by protease 1 after residue -9 or - 10 (generating an eight- or nine-residue intermediate) and then by protease 11. mtps with Arg in position -2 but not at - 10 are likely to be cleaved only by protease I. Good candidates for two-step processing would thus be, for example, bovine adrenodoxin reductase, cytochrome oxidase IV and proteolipid PI and human pyruvate dehydrogenase b-subunit. The yeast cytochrome oxidase IV mtp and the Neurospora Rieske Fe-S and ATPase-subunit-9 mtps are also cleaved in two steps. The cytochrome oxidase IV mtp is cleaved between positions - 9 and - 8 (Arg-ThrlLeu) by the matrix protease isolated from yeast and cleaved at the final cleavage site (Leu- LeulGln) upon import into intact mitochondria [26]. When the final site is deleted, only the first cleavage takes place upon import. The Rieske Fe-S mtp is also cleaved first between positions - 9 and - 8 (Arg-AlaJLeu) and then at the final site (Leu-GlnlGly) [27]. Both mtps thus have Arg in position - 10 followed by an apolar Leu in position - 8 and neither has Arg in position -2 (it should be noted, though, that the second cleavage of the Rieske Fe-S mtp might be related to translocation of the protein back through the inner membrane, and that it is not known whether this cleavage takes place in the matrix or the intermembrane space). The ATPasesubunit-9 mtp, on the other hand, is cleaved initially between residues - 32 and ~ 31 (Arg-ThrJIle) and then at the final site (Arg-AlajTyr) [28]; in this case it would seem that protease I is responsible for both cuts. Whether two distinct proteases exist in Neurospora and yeast is unknown but seems likely in view of these results. In summary, part of the protease-i-recognition sequence seems to be an Arg in position - 2 relative to the cleavage site. Protease I1 seems to have the peculiar property of removing precisely eight or nine residues from the amino terminus of a precursor protein; it seems to be an octapeptidyl peptidase. The reason(s) for two-step processing of some mtps can only be speculated upon; an attractive idea is that the short extension on the intermediate may be required for correct folding of the mature protein, or for its assembly into larger multisubunit complexes. Whether multiple cleavage enzymes can take part in the processing of stroma-targeting ctps is not known. If proper folding or assembly of the mature protein can be controlled by amino-terminal extensions, chloroplasts may also have envolved multistep processing of targeting peptides. Processing intermediates have been found for pea ribulose bisphosphate carboxylase/oxygenase small subunit [29, 301 and for the L18 ribosomal protein from Chlamydomonas [31], but the intermediate cleavage sites were not determined. The plastid-encoded CFo-I and CFo-IV subunits are synthesized with polar amino-terminal extensions of 17 and 18 residues, respectively [32, 331. It is thus not unlikely that chloroplasts will be found to be similar to mitochondria in this respect. Intra-organellar sorting signals: remnants of a secretory pathway? The remarkable similarity between the intra-organellar sorting signals (targeting proteins to the mitochondria1 intermeinbrane space and the thylakoid lumen) and secretory signal peptides is a strong indication that the secretory machinery of the bacterial precursors to mitochondria and chloroplasts has been retained in the present day organelles. Indeed, the cleavage sites in the precursors of proteins of the thylakoid lumen are of the same type as in bacterial signal peptides (A-X-AJ), whereas only some of the cleavage sites in precursors of intermembrane-space proteins follow this pattern. There is a marked gradation in the overall amino acid composition of the apolar regions in the eukaryotic, prokaryotic and intra-organellar signal peptides (Fig. 5). Although the content of Ala + Leu remains fairly constant, eukaryotic signal peptides have a strong preference for Leu, prokaryotic signal peptides contain equal numbers of Ala and Leu residues and the intra-organellar signals have a strong bias towards Ala. Thus, although similar in basic design, eukaryotic signal peptides are markedly more hydrophobic than the intra-organellar sorting signals. It is conceivable that the latter could be erroneously recognized as secretory signal peptides if they contained apolar regions of a mean hydrophobicity comparable to that of true eukaryotic signal peptides [34]. In line with this speculation, the apolar region of the thylakoid-targeting signal peptide of the chloroplastencoded cytochromef protein contains no Ala but many Leu, Ile and Val residues (Fig. 4). Since this protein is made inside the chloroplast, it obviously cannot be mistakenly routed into the secretory pathway. Finally, we note that if mosaic ctps of this kind route proteins to the thylakoid lumen, some other kind of targeting signal must be used for routing to the interenvelope space PSI. This work was supported by a grant from the Swedish Natural Sciences Research Council to GvH. REFERENCES 1. Roise, D., Horvath, S. J., Tomich, J. M., Richards, J. H. & Schatz, G. (1986) EMBO J. 5, von Heijne, G. (1986) EMBO J. 5, Roise, D., Theiler, F., Horvath, S. J., Tomich, J. M., Richards, J. H., Allison, D. S. & Schatz, G. (1988) EMBO 1. 7, Karlin-Neumann, G. A. & Tobin, E. M. (1986) EMBO J. 5, Schmidt, C. W. & Mishkind, M. L. (1986) Annu. Rev. Biochem. 55, Wasmann, C. C., Reiss, B. & Bohncrt, H. J. (1988) J. Bid. Chem. 263, Eisenberg, D., Weiss, R. M. & Terwilliger, T. C. (1984) Proc. Nut1 Acud. Sci. USA 81, Tyagi, A., Hermans, J., Steppuhn, J., Janson, C., Vater, F. & Herrmann, R. G. (1987) Mol. & Gen. Genet. 207, Levitt, M. (1978) Biochemistry 17, Hartl, F., Ostermann, J., Guiard, B. & Neupert, W. (1987) Cell 51,
10 11. Smeekens, S., Bauerle, C., Hageman, J., Kecgstra, K. & Weisbeek, P. (1986) Cell46, von Heijne, G. (1985) J. Mol. Biol. 184, Chothia, C. (1984) Annu. Rev. Biochem. 53, Flinta, C., Persson, B., Jornvall, H. & von Heijne, G. (1986) Eur. J. Biochem. 154, Huang, S., Elliott, R. C., Liu, P. S., Koduri, R. K., Weickmann, J. L., Lee, J. H., Blair, L. C., Ghosh-Dastidar, P., Bradshaw, R. A., Bryan, K. M., Einarson, B., Kendall, R. L., Kolacz, K. H. & Saito, K. (1988) Biochemistry 26, Rciss, B., Wasmann, C. C. & Bohnert, H..I. (1987) Mol. & Gen. Genet. 209, Tamm, L. (1986) Biochemistry 25, Eilers, M. & Schatz, G. (1988) Cell 52, Endo, T. & Schatz, G. (1988) EMBO J. 7, Ou, W.-J., Ito, A,, Umeda, M., Inoue, K. & Omura, T. (1988) J. Biochem. (Tokyo) 103, Mcrchant, S. & Bogorad, L. (1987) J. Biol. Chem. 262, Miura, S., Amaya, Y. & Mori, M. (1Y86) Biochem. Biophys. Res. Commun. 134, Kalousek, F., Hendrick, J. P. & Rosenbcrg, L. E. (1988) Proc. Nutl Acad. Sci. USA, in the press. 24. Sztul, E. S., Hendrick, J. P., Kraus, J. P., Wall, D., Kalousek, F. & Rosenberg, L. E. (1987) J. Cell Biol. 105, Sztul, E. S., Chu, T. W., Strauss, A. W. & Rosenberg, L. E. (1988) J. Biol. Chem. 263, Hurt, E. C., Peshold-Hurt, B., Suda, K., Opplinger, W. & Schatz, G. (1985) EMBO J. 4, Hartl, F., Schmidt, B., Wachter, E., Weiss, H. & Neupert, W. (1986) Cell47, Schmidt, B., Wachter, E., Sebald, W. & Neupert, W. (1984) Eur. J. Biochem. 144, Robinson, C. & Ellis, R. J. (1984) Eur. J. Biochem. 142, Mishkind, M. L., Wcssler, S. R. & Schmidt, G. W. (1985) J. Cell Bid. 100, Schmidt, R. J., Gillham, N. W. & Boynton, J. E. (1985) Mol. Cell Biol. 5, Bird, C. R., Koller, B., Auffrct, A. D., Huttly, A. K., Howe, C. J., Dyer, T. A. &Gray, J. C. (1985) EMBO J. 4, Hermans, J., Rother, C., Bichler, J., Steppuhn, J. & Herrmann, R. G. (1988) Plant Mol. Bid. 10, von Heijnc, G. (1986) FEBS Lett. 198, 1-4. Soll, J. & Bennett, J. (1988) Eur. J. Biochem. 175, von Hcijne, G. (1987) Protein Sequence and Dato Analysis 1, Eiscnberg, D. (1984) Annu. Rev. Biochem. 53, Chen, W.-J. & Douglas, M. G. (1987) J. Biol. Chem.262, Velours, J., Durrens, P., Aigle, M. & Guerin, B. (1988) Eur. J. Biochem. 170, Maarse, A. C., Van Loon, A. P. G. M., Riezman, H., Gregor, I., Schatz, G. & Grivell, L. A. (1984) EMBO J. 3, Seraphin, B., Simon, M. & Faye, G. (1985) Curr. Genet. 9, Wright, R. M., KO, C., Cumsky, M. G. & Poyton, R. 0. (1984) J. Biol. Chem. 259, Patterson, T. E. & Poyton, R. 0. (1986) J. Biol. Chem. 261, Marres, C. A. M., Van Loon, A. P. G. M., Oudshoorn, P., Van Steeg, H., Grivell, L. A. & Slater, E. C. (1985) Eur. J. Biochem. 147, Beckmann, J. D., Ljungdahl, P. O., Lopez, J. L. & Trumpower, B. L. (1987) J. Bid. Chem. 262, Shannon, K. W. & Rabinowitz, J. C. (1988) J. Bid. Chem. 263, Browning, K. S., Uhlinger, D. J. & Reed, L. J. (1988) Proc. Nut/ A~ad. Sci. USA 8.5, Sachs, M. S., David, M., Werner, S. & RajBhandary, U. L. (1986) J. Biol. Chem. 261, Viebrock, A., Perz, A. & Sebald, W. (1982) EMBO J. I, Okamura, T., John, M. E., Zuber, M. X., Simpson, E. R. & Waterman, M. R. (1985) Proc. Natl Acad. Sci. USA 82, Sagara, Y., Takata, Y., Miyata, T., Hara, T. & Horiuchi, T. (1987) J. Biochem. (Tokyo) 102, Walker, J. E., Gay, N. J., Powell, S. J., Kostina, M. & Dyer, M. R. (1987) Biochemistry 26, Lomax, M. I., Bachman, N. J., Nasoff, M. S., Caruthers, M. H. & Grossman, L. I. (1984) Proc. Nutl Acad. Sci. USA 81, Morohashi, K., Fujii-Kuriyama, Y., Okada, Y., Sogawa, K., Hirose, T., Inayama, S. & Omura, T. (1984) Proc. Nut1 Acad. Sci. USA 81, Morohashi, K., Yoshioka, H., Gotoh, O., Okada, Y., Yamamoto, K., Miyata, T., Sogawa, K., Fujii-Kuriyama, Y. & Omura, T. (1987) J. Biochem. (Tokyo) 102, Gay, N. J. & Walker, J. E. (1985) EMBO J. 4, Brothwick, I. A., Srivastava, G., Day, A. R., Pirola, B. A,, Snoswcll, M. A., May, B. K. & Elliott, W. H. (1985) Eur. J. Biochem. 150, Jaussi, R., Cotton, B., Juretic, N., Christen, P. & Schiimperli, D. (1985) J. Biol. Chem. 260, Ohta, S. & Kagawa, Y. (1986) J. Biochem. (Tokyo) 99, Otulakowski, G. & Robinson, B. H. (1987) J. Bid. Chem. 262, Ho, Y.3. & Crapo, J. D. (1988) FEBS Lett. 229, Horwich, A,, Fenton, W. A., Williams, K. R., Kalousek, F., Kraus, J. P., Doolittle, R. F., Konigsberg, W. & Rosenberg, L. E. (1984) Science (Wash. DC) 224, Koike, K., Ohta, S., Urata, Y., Kagawa, Y. & Koike, M. (1988) Proc. Natl Acad. Sci. USA 8.5, Nyunoya, H., Broglic, K. E., Widgren, E. E. & Lusty, C. J. (1985) J. Biol. Chem. 260, Grant, P. M., Tellam, J., May, V. L. & Strauss, A. W. (1986) Nucleic Acids Res. 14, Oda, T., Miyajima, H., Suzuki, Y. & Ichiyama, A. (1987) Eur. J. Biochem. 168, Henning, W. D., Upton, C., McFadden, G., Majumdar, R. & Bridger, W. A. (1988) Proc. Nut1 Acud. Sci. USA 85, Farrcs, J., Guan, K.-L. & Weiner, H. (1988) Biochem. Biophys. Rex Commun. 150, Lautner, A., Klein, R., Ljungberg, U., Reilander, H., Bartling, D., Andersson, B., Reinke, H., Bcyreuther, K. & Hermann, R. G. (1988).I. Riol. Chem. 263, Jansen, T., Keilindcr, H., Steppuhn, J. & Herrmann, R. G. (1988) Curr. Genet. 13, Steppuhn, J., Rother, C., Hermans, J., Jansen, T., Salnikow, J., Hauska, G. & Herrmann, R. G. (1987) Mol. & Gen. Genet. 210, Tittgen, J. (1985) Thesis, University of Diisseldorf. 73. Werneke, J. M., Zielinski, R. E. & Ogren, W. L. (1988) Proc. Nail Acad. Sci. USA 85, Back, E., Burkhart, W., Moyer, M., Privalle, L. & Rothstein, S. (1988) Mol. & Gen. Genet. 212, Scherer, D. E. & Knauf, V. C. (1987) Plant Mol. Bid. 9, Steppuhn, J., Hermans, J., Nechustai, R., Ljungberg, U., Thiimmler, F., Lottspeich, F. & Herrmann, R. G. (1988) FEBS Lett. 237, Smeekens, S., van Binsbergen, J. & Weisbeek, P. (1985) Nucleic Acids Res. 13, Kolanus, W., Scharnhost, C., Kuhne, U. & Herzfeld. F. (1987) Mol. & Gen. Genet. 209, Vierling, E., Nagao, R. T., DeRocher, A. E. & Harris, L. M. (1988) EMBO J. 7, Tingey, S. V., Walker, E. L. & Coruzzi, G. M. (1987) EMBO J. 6, 1-9.
11 Hoffman, N. E., Pichersky, E., Malik, V. S., KO, K. & Cashmore, A. R. (1988) Plant Mol. Bid. 10, Pichersky, E., Bernatzky, R., Tanksley, S. D., Breidenbach, R. B., Kausch, A. P. & Cashmore, A. R. (1985) Gene (Amst.) 40, Hoffman, N. E., Pichersky, E., Malik, V. S., Castresana, C., KO, K., Dam, S. C. & Cashinore, A. R. (1987) Proc. Natl Acad. Sci. USA 84, Rose, R. E., DeJesus, C. E., Moylan, S. L., Ridge, N. P., Scherer, D. E. & Knauf, V. C. (1987) Nucleic Acids Res. 15, Schmidt, G. W., Devillers-Thiery, A,, Desrusseaux, H., Blobel, G. & Chua, N.-H. (1979) J. Cell Biol. 83, Klee, H. J., Muskopf, Y. M. & Gasscr, C. S. (1987) Mol. & Gen. Genet. 210, Sadler, I., Suda, K., Schatz, G., Kaudewitz, F. & Haid, A. (3 984) EMBO J. 3, Guiard, B. (1985) EMBO J. 4, Kaput, J., Goltz, S. & Blobel, G. (1982) J. Biol. Chern. 257, Nishikimi, M., Ohta, S., Suzuki, H., Tanaka, T., Kikkawa, F., Tanaka, M., Kagawa, Y. & Ozawa, T. (1988) Nucleic Acids Res. 16, Jansen, T., Rother, C., Steppuhn, J., Reinke, H., Beyreuther, K., Jansson, C., Anderson, B. & Herrmanii, R. G. (1987) FEBS Lett. 216, Rother, C., Jansen, T., Tyagi, A,, Tiltgcn, J. & Herrmann, R. G. (1986) Curr. Genet. 11, Mayfield, S. P., Rahre, M., Frank, G., Zuber, H. & Rochaix, J.-D. (1987) Proc. Natl Acad. Sci. USA 84, Willey, D. L., Auffret, A. D. &Gray, J. C. (1984) CeN36, Akashi, A,, Yoshida, Y., Nakagoshi, H., Kuroki, K., Hashimoto, T., Tagawa, K. & Imamoto, F. (1988) J. Biochem. (Tokyo) 104, White, J. A. & Scandalios, J. G. (1988) Biochim. Biophys. Actu 951,61-70.
Isolda Purchase - EDI
Isolda Purchase - EDI Document v 1.0 1 Table of Contents Table of Contents... 2 1 Introduction... 3 1.1 What is EDI?... 4 1.2 Sending and receiving documents... 4 1.3 File format... 4 1.3.1 XML (language
Läs merThis exam consists of four problems. The maximum sum of points is 20. The marks 3, 4 and 5 require a minimum
Examiner Linus Carlsson 016-01-07 3 hours In English Exam (TEN) Probability theory and statistical inference MAA137 Aids: Collection of Formulas, Concepts and Tables Pocket calculator This exam consists
Läs merIsometries of the plane
Isometries of the plane Mikael Forsberg August 23, 2011 Abstract Här följer del av ett dokument om Tesselering som jag skrivit för en annan kurs. Denna del handlar om isometrier och innehåller bevis för
Läs merLabokha AA et al. xlnup214 FG-like-1 xlnup214 FG-like-2 xlnup214 FG FGFG FGFG FGFG FGFG xtnup153 FG FGFG xtnup153 FG xlnup62 FG xlnup54 FG FGFG
xlnup214 FG-like-1 (aa 443-69) TSVSAPAPPASAAPRSAAPPPYPFGLSTASSGAPTPVLNPPASLAPAATPTKTTSQPAAAATSIFQPAGPAAGSLQPPSLPAFSFSSANNAANASAPSSFPFGA AMVSSNTAKVSAPPAMSFQPAMGTRPFSLATPVTVQAATAPGFTPTPSTVKVNLKDKFNASDTPPPATISSAAALSFTPTSKPNATVPVKSQPTVIPSQASVQP
Läs merBiochemistry 201 Advanced Molecular Biology (
Biochemistry 201 Advanced Molecular Biology (http://cmgm cmgm.stanford.edu/biochem201/) Bioinformatics: Discovering Function from Sequence Doug Brutlag Departments of Biochemistry June 4, 1999 Discovering
Läs merRättningstiden är i normalfall 15 arbetsdagar, annars är det detta datum som gäller:
Molekylärbiologi Provmoment: Ladokkod: Tentamen ges för: Tentamen TK151C Bt3 7,5 högskolepoäng TentamensKod: Tentamensdatum: 2016-01-12 Tid: 14:00 18:00 Hjälpmedel: Tillåtna hjälpmedel är lexikon. Dock
Läs merStiftelsen Allmänna Barnhuset KARLSTADS UNIVERSITET
Stiftelsen Allmänna Barnhuset KARLSTADS UNIVERSITET National Swedish parental studies using the same methodology have been performed in 1980, 2000, 2006 and 2011 (current study). In 1980 and 2000 the studies
Läs merFORSKNINGSKOMMUNIKATION OCH PUBLICERINGS- MÖNSTER INOM UTBILDNINGSVETENSKAP
FORSKNINGSKOMMUNIKATION OCH PUBLICERINGS- MÖNSTER INOM UTBILDNINGSVETENSKAP En studie av svensk utbildningsvetenskaplig forskning vid tre lärosäten VETENSKAPSRÅDETS RAPPORTSERIE 10:2010 Forskningskommunikation
Läs merRastercell. Digital Rastrering. AM & FM Raster. Rastercell. AM & FM Raster. Sasan Gooran (VT 2007) Rastrering. Rastercell. Konventionellt, AM
Rastercell Digital Rastrering Hybridraster, Rastervinkel, Rotation av digitala bilder, AM/FM rastrering Sasan Gooran (VT 2007) Önskat mått * 2* rastertätheten = inläsningsupplösning originalets mått 2
Läs mer8 < x 1 + x 2 x 3 = 1, x 1 +2x 2 + x 4 = 0, x 1 +2x 3 + x 4 = 2. x 1 2x 12 1A är inverterbar, och bestäm i så fall dess invers.
MÄLARDALENS HÖGSKOLA Akademin för utbildning, kultur och kommunikation Avdelningen för tillämpad matematik Examinator: Erik Darpö TENTAMEN I MATEMATIK MAA150 Vektoralgebra TEN1 Datum: 9januari2015 Skrivtid:
Läs merGrafisk teknik IMCDP IMCDP IMCDP. IMCDP(filter) Sasan Gooran (HT 2006) Assumptions:
IMCDP Grafisk teknik The impact of the placed dot is fed back to the original image by a filter Original Image Binary Image Sasan Gooran (HT 2006) The next dot is placed where the modified image has its
Läs merStyrteknik: Binära tal, talsystem och koder D3:1
Styrteknik: Binära tal, talsystem och koder D3:1 Digitala kursmoment D1 Boolesk algebra D2 Grundläggande logiska funktioner D3 Binära tal, talsystem och koder Styrteknik :Binära tal, talsystem och koder
Läs mer1. Compute the following matrix: (2 p) 2. Compute the determinant of the following matrix: (2 p)
UMEÅ UNIVERSITY Department of Mathematics and Mathematical Statistics Pre-exam in mathematics Linear algebra 2012-02-07 1. Compute the following matrix: (2 p 3 1 2 3 2 2 7 ( 4 3 5 2 2. Compute the determinant
Läs merResultat av den utökade första planeringsövningen inför RRC september 2005
Resultat av den utökade första planeringsövningen inför RRC-06 23 september 2005 Resultat av utökad första planeringsövning - Tillägg av ytterligare administrativa deklarationer - Variant (av case 4) med
Läs merSUPPLEMENTARY FIGURE LEGENDS
SUPPLEMETARY FIGURE LEGEDS Supplementary Fig. 1. Flow cytometric analysis of wildtype, mutant and chimeric protein surface expression. Cells transduced with the individual constructs indicated were stained
Läs merRoom E3607 Protein bioinformatics Protein Bioinformatics. Computer lab Tuesday, May 17, 2005 Sean Prigge Jonathan Pevsner Ingo Ruczinski
Room E3607 Protein bioinformatics 260.841 Protein Bioinformatics Computer lab Tuesday, May 17, 2005 Sean Prigge Jonathan Pevsner Ingo Ruczinski Outline of today s lab Topic Suggested time 1 Find a protein
Läs merAborter i Sverige 2008 januari juni
HÄLSA OCH SJUKDOMAR 2008:9 Aborter i Sverige 2008 januari juni Preliminär sammanställning SVERIGES OFFICIELLA STATISTIK Statistik Hälsa och Sjukdomar Aborter i Sverige 2008 januari juni Preliminär sammanställning
Läs merWriting with context. Att skriva med sammanhang
Writing with context Att skriva med sammanhang What makes a piece of writing easy and interesting to read? Discuss in pairs and write down one word (in English or Swedish) to express your opinion http://korta.nu/sust(answer
Läs merUttagning för D21E och H21E
Uttagning för D21E och H21E Anmälan till seniorelitklasserna vid O-Ringen i Kolmården 2019 är öppen fram till och med fredag 19 juli klockan 12.00. 80 deltagare per klass tas ut. En rangordningslista med
Läs merLUNDS TEKNISKA HÖGSKOLA Institutionen för Elektro- och Informationsteknik
LUNDS TEKNISKA HÖGSKOLA Institutionen för Elektro- och Informationsteknik SIGNALBEHANDLING I MULTIMEDIA, EITA50, LP4, 209 Inlämningsuppgift av 2, Assignment out of 2 Inlämningstid: Lämnas in senast kl
Läs merGrafisk teknik IMCDP. Sasan Gooran (HT 2006) Assumptions:
Grafisk teknik Sasan Gooran (HT 2006) Iterative Method Controlling Dot Placement (IMCDP) Assumptions: The original continuous-tone image is scaled between 0 and 1 0 and 1 represent white and black respectively
Läs merFysisk aktivitet och hjärnan
1 Fysisk aktivitet och hjärnan Professor Ingibjörg H. Jónsdóttir Hälsan och stressmedicin, VGR Institutionen för kost och idrottsvetenskap Göteborgs Universitet Kvinnlig simultankapacitet troligen en myt
Läs merGrafisk teknik. Sasan Gooran (HT 2006)
Grafisk teknik Sasan Gooran (HT 2006) Iterative Method Controlling Dot Placement (IMCDP) Assumptions: The original continuous-tone image is scaled between 0 and 1 0 and 1 represent white and black respectively
Läs merÖvning 5 ETS052 Datorkommuniktion Routing och Networking
Övning 5 TS5 Datorkommuniktion - 4 Routing och Networking October 7, 4 Uppgift. Rita hur ett paket som skickas ut i nätet nedan från nod, med flooding, sprider sig genom nätet om hop count = 3. Solution.
Läs merEn bild säger mer än tusen ord?
Faculteit Letteren en Wijsbegeerte Academiejaar 2009-2010 En bild säger mer än tusen ord? En studie om dialogen mellan illustrationer och text i Tiina Nunnallys engelska översättning av Pippi Långstrump
Läs merKurskod: TAMS28 MATEMATISK STATISTIK Provkod: TEN1 05 June 2017, 14:00-18:00. English Version
Kurskod: TAMS28 MATEMATISK STATISTIK Provkod: TEN1 5 June 217, 14:-18: Examiner: Zhenxia Liu (Tel: 7 89528). Please answer in ENGLISH if you can. a. You are allowed to use a calculator, the formula and
Läs merViktig information för transmittrar med option /A1 Gold-Plated Diaphragm
Viktig information för transmittrar med option /A1 Gold-Plated Diaphragm Guldplätering kan aldrig helt stoppa genomträngningen av vätgas, men den får processen att gå långsammare. En tjock guldplätering
Läs mer- den bredaste guiden om Mallorca på svenska! -
- den bredaste guiden om Mallorca på svenska! - Driver du företag, har en affärsrörelse på Mallorca eller relaterad till Mallorca och vill nå ut till våra läsare? Då har du möjlighet att annonsera på Mallorcaguide.se
Läs merPORTSECURITY IN SÖLVESBORG
PORTSECURITY IN SÖLVESBORG Kontaktlista i skyddsfrågor / List of contacts in security matters Skyddschef/PFSO Tord Berg Phone: +46 456 422 44. Mobile: +46 705 82 32 11 Fax: +46 456 104 37. E-mail: tord.berg@sbgport.com
Läs merPreschool Kindergarten
Preschool Kindergarten Objectives CCSS Reading: Foundational Skills RF.K.1.D: Recognize and name all upper- and lowercase letters of the alphabet. RF.K.3.A: Demonstrate basic knowledge of one-toone letter-sound
Läs merKurskod: TAIU06 MATEMATISK STATISTIK Provkod: TENA 15 August 2016, 8:00-12:00. English Version
Kurskod: TAIU06 MATEMATISK STATISTIK Provkod: TENA 15 August 2016, 8:00-12:00 Examiner: Xiangfeng Yang (Tel: 070 0896661). Please answer in ENGLISH if you can. a. Allowed to use: a calculator, Formelsamling
Läs merCHANGE WITH THE BRAIN IN MIND. Frukostseminarium 11 oktober 2018
CHANGE WITH THE BRAIN IN MIND Frukostseminarium 11 oktober 2018 EGNA FÖRÄNDRINGAR ü Fundera på ett par förändringar du drivit eller varit del av ü De som gått bra och det som gått dåligt. Vi pratar om
Läs merValidering av kvalitetsregisterdata vad duger data till?
Validering av kvalitetsregisterdata vad duger data till? Anders Ekbom, Professor Karolinska Institutet Institutionen för medicin Solna Enheten för klinisk epidemiologi Karolinska Universitetssjukhuset
Läs merF ξ (x) = f(y, x)dydx = 1. We say that a random variable ξ has a distribution F (x), if. F (x) =
Problems for the Basic Course in Probability (Fall 00) Discrete Probability. Die A has 4 red and white faces, whereas die B has red and 4 white faces. A fair coin is flipped once. If it lands on heads,
Läs merModule 6: Integrals and applications
Department of Mathematics SF65 Calculus Year 5/6 Module 6: Integrals and applications Sections 6. and 6.5 and Chapter 7 in Calculus by Adams and Essex. Three lectures, two tutorials and one seminar. Important
Läs mer12.6 Heat equation, Wave equation
12.6 Heat equation, 12.2-3 Wave equation Eugenia Malinnikova, NTNU September 26, 2017 1 Heat equation in higher dimensions The heat equation in higher dimensions (two or three) is u t ( = c 2 2 ) u x 2
Läs merNormalfördelning. Modeller Vi har alla stött på modeller i olika sammanhang. Ex:
Normalfördelning 1 Modeller Vi har alla stött på modeller i olika sammanhang. Ex: Leksaksbilar Modelljärnvägar Dockskåp 2 En leksaksbil är i vissa avseenden en kopia av en riktig bil. Men den skiljer sig
Läs merSUZUKI GRAND VITARA 3P CITY 2011»
00-2082 120 SUZUKI GRAND VITARA 3P CITY 2011» 661-0830 rev. 2014-04-04 DC Congratulations on purchasing an ATS towbar Alexo Towbars Sweden offer quality towbars produced as a result of direct market research.
Läs merÖvning 4 EITF25 & EITF Protokoll. October 29, 2016
- 2016 Protokoll October 29, 2016 1 Uppgift 1. Nedan finns en Ethernet II-ram där Preamble, SFD och CRC är borttagna. Ramen är beskriven i hexadecimalt format. Svara på följande frågor genom att studera
Läs merFÖRBERED UNDERLAG FÖR BEDÖMNING SÅ HÄR
FÖRBERED UNDERLAG FÖR BEDÖMNING SÅ HÄR Kontrollera vilka kurser du vill söka under utbytet. Fyll i Basis for nomination for exchange studies i samråd med din lärare. För att läraren ska kunna göra en korrekt
Läs merKönsfördelningen inom kataraktkirurgin. Mats Lundström
Könsfördelningen inom kataraktkirurgin Mats Lundström Innehåll Fördelning av antal operationer utveckling Skillnader i väntetid Effekt av NIKE Skillnader i synskärpa före operation Skillnader i Catquest-9SF
Läs merModule 1: Functions, Limits, Continuity
Department of mathematics SF1625 Calculus 1 Year 2015/2016 Module 1: Functions, Limits, Continuity This module includes Chapter P and 1 from Calculus by Adams and Essex and is taught in three lectures,
Läs merA QUEST FOR MISSING PULSARS
LOFAR A QUEST FOR MISSING PULSARS Samayra Straal Joeri v. Leeuwen WHAT ARE MISSING ~ half of PWN are associated with a pulsar (32/56) PULSARS? less than 25% of all SNRs are associated with a pulsar (60/294)
Läs merA study of the performance
A study of the performance and utilization of the Swedish railway network Anders Lindfeldt Royal Institute of Technology 2011-02-03 Introduction The load on the railway network increases steadily, and
Läs merThe Arctic boundary layer
The Arctic boundary layer Interactions with the surface, and clouds, as learned from observations (and some modeling) Michael Tjernström Department of Meteorology & the Bert Bolin Center for Climate Research,
Läs merMake a speech. How to make the perfect speech. söndag 6 oktober 13
Make a speech How to make the perfect speech FOPPA FOPPA Finding FOPPA Finding Organizing FOPPA Finding Organizing Phrasing FOPPA Finding Organizing Phrasing Preparing FOPPA Finding Organizing Phrasing
Läs mer2 Uppgifter. Uppgifter. Svaren börjar på sidan 35. Uppgift 1. Steg 1. Problem 1 : 2. Problem 1 : 3
1 2 Uppgifter Uppgifter Svaren börjar på sidan 35. Uppgift 1. Steg 1 Problem 1 : 2 Problem 1 : 3 Uppgifter 3 Svarsalternativ. Answer alternative 1. a Svarsalternativ. Answer alternative 1. b Svarsalternativ.
Läs merREHAB BACKGROUND TO REMEMBER AND CONSIDER
Training in water REHAB BACKGROUND TO REMEMBER AND CONSIDER PHASE I: PROLIFERATION PROTECTION, 0-6 WEEKS PHASE II: TRANSITION PROGRESSION, 7-12 WEEKS PHASE III: REMODELLING FUNCTION, 13-32 WEEKS PHASE
Läs merWindlass Control Panel v1.0.1
SIDE-POWER Windlass Systems 86-08950 Windlass Control Panel v1.0.1 EN Installation manual Behåll denna manual ombord! S Installations manual SLEIPNER AB Kilegatan 1 452 33 Strömstad Sverige Tel: +46 525
Läs merDokumentnamn Order and safety regulations for Hässleholms Kretsloppscenter. Godkänd/ansvarig Gunilla Holmberg. Kretsloppscenter
1(5) The speed through the entire area is 30 km/h, unless otherwise indicated. Beware of crossing vehicles! Traffic signs, guardrails and exclusions shall be observed and followed. Smoking is prohibited
Läs merEXTERNAL ASSESSMENT SAMPLE TASKS SWEDISH BREAKTHROUGH LSPSWEB/0Y09
EXTENAL ASSESSENT SAPLE TASKS SWEDISH BEAKTHOUGH LSPSWEB/0Y09 Asset Languages External Assessment Sample Tasks Breakthrough Stage Listening and eading Swedish Contents Page Introduction 2 Listening Sample
Läs merMichael Q. Jones & Matt B. Pedersen University of Nevada Las Vegas
Michael Q. Jones & Matt B. Pedersen University of Nevada Las Vegas The Distributed Application Debugger is a debugging tool for parallel programs Targets the MPI platform Runs remotley even on private
Läs merConsumer attitudes regarding durability and labelling
Consumer attitudes regarding durability and labelling 27 april 2017 Gardemoen Louise Ungerth Konsumentföreningen Stockholm/ The Stockholm Consumer Cooperative Society louise.u@konsumentforeningenstockholm.se
Läs merIntroduktion till vetenskaplig metodik. Johan Åberg
Introduktion till vetenskaplig metodik Johan Åberg Innehåll Forskarvärlden Viktiga begrepp Referenshantering Den vetenskapliga rapporten Vetenskaplig diskussion Forskarvärlden Forskare mäts i antal publikationer
Läs merSupport Manual HoistLocatel Electronic Locks
Support Manual HoistLocatel Electronic Locks 1. S70, Create a Terminating Card for Cards Terminating Card 2. Select the card you want to block, look among Card No. Then click on the single arrow pointing
Läs merPRESS FÄLLKONSTRUKTION FOLDING INSTRUCTIONS
PRESS FÄLLKONSTRUKTION FOLDING INSTRUCTIONS Vänd bordet upp och ner eller ställ det på långsidan. Tryck ner vid PRESS och fäll benen samtidigt. OBS! INGA STORA KRAFTER KRÄVS!! Om benen sitter i spänn tryck
Läs merAccomodations at Anfasteröd Gårdsvik, Ljungskile
Accomodations at Anfasteröd Gårdsvik, Ljungskile Anfasteröd Gårdsvik is a campsite and resort, located right by the sea and at the edge of the forest, south west of Ljungskile. We offer many sorts of accommodations
Läs merThe Finite Element Method, FHL064
The Finite Element Method, FHL064 Division of Solid Mechanics Course program, vt2, 20 Course description The finite element method (FEM) is a numerical method able to solve differential equations, i.e.
Läs merWebbregistrering pa kurs och termin
Webbregistrering pa kurs och termin 1. Du loggar in på www.kth.se via den personliga menyn Under fliken Kurser och under fliken Program finns på höger sida en länk till Studieöversiktssidan. På den sidan
Läs merDocumentation SN 3102
This document has been created by AHDS History and is based on information supplied by the depositor /////////////////////////////////////////////////////////// THE EUROPEAN STATE FINANCE DATABASE (Director:
Läs merCustom-made software solutions for increased transport quality and creation of cargo specific lashing protocols.
Custom-made software solutions for increased transport quality and creation of cargo specific lashing protocols. ExcelLoad simulates the maximum forces that may appear during a transport no matter if the
Läs merTentamen Biokemi 2 KEM090
Tentamen Biokemi 2 KEM090 2011 10 28 Max: 70 poäng Godkänt: 35 poäng Väl godkänt: 52 poäng Inga hjälpmedel tillåtna OBS! Besvara inte mer än en fråga per sida! Markera varje sida med personlig kod, datum
Läs merEXPERT SURVEY OF THE NEWS MEDIA
EXPERT SURVEY OF THE NEWS MEDIA THE SHORENSTEIN CENTER ON THE PRESS, POLITICS & PUBLIC POLICY JOHN F. KENNEDY SCHOOL OF GOVERNMENT, HARVARD UNIVERSITY, CAMBRIDGE, MA 0238 PIPPA_NORRIS@HARVARD.EDU. FAX:
Läs merTentamen Molekylärbiologi X3 (1MB608) 10 March, 2008 Page 1 of 5. Skriv svaren på varje fråga på SEPARATA blad.
Tentamen Molekylärbiologi X3 (1MB608) 10 March, 2008 Page 1 of 5 Skriv svaren på varje fråga på SEPARATA blad. Skriv namn på VARJE blad. Du kan svara på engelska eller svenska. Motivera eller förklara
Läs merSkill-mix innovation in the Netherlands. dr. Marieke Kroezen Erasmus University Medical Centre, the Netherlands
Skill-mix innovation in the Netherlands dr. Marieke Kroezen Erasmus University Medical Centre, the Netherlands m.kroezen@erasmusmc.nl The skill-mix innovation of interest BEFORE AFTER How did the Netherlands
Läs merHållbar utveckling i kurser lå 16-17
Hållbar utveckling i kurser lå 16-17 : Jag tillhör akademin / My position is in the School of Jag tillhör akademin / My position is in the School of Humaniora och medier / Humanities and Media Studies
Läs merTheory 1. Summer Term 2010
Theory 1 Summer Term 2010 Robert Elsässer 1 Introduction Summer Term 2010 Robert Elsässer Prerequisite of Theory I Programming language, such as C++ Basic knowledge on data structures and algorithms, mathematics
Läs merExam Molecular Bioinformatics X3 (1MB330) - 1 March, Page 1 of 6. Skriv svar på varje uppgift på separata blad. Lycka till!!
Exam Molecular Bioinformatics X (MB) - March, - Page of Skriv svar på varje uppgift på separata blad. Lycka till!! Write the answers to each of the questions on separate sheets of paper. ood luck!! ) Sequence
Läs merCUSTOMER READERSHIP HARRODS MAGAZINE CUSTOMER OVERVIEW. 63% of Harrods Magazine readers are mostly interested in reading about beauty
79% of the division trade is generated by Harrods Rewards customers 30% of our Beauty clients are millennials 42% of our trade comes from tax-free customers 73% of the department base is female Source:
Läs merInstallation Instructions
Installation Instructions (Cat. No. 1794-IE8 Series B) This module mounts on a 1794 terminal base unit. 1. Rotate keyswitch (1) on terminal base unit (2) clockwise to position 3 as required for this type
Läs merAlias 1.0 Rollbaserad inloggning
Alias 1.0 Rollbaserad inloggning Alias 1.0 Rollbaserad inloggning Magnus Bergqvist Tekniskt Säljstöd Magnus.Bergqvist@msb.se 072-502 09 56 Alias 1.0 Rollbaserad inloggning Funktionen Förutsättningar Funktionen
Läs merSAMMANFATTNING AV SKILLNADER MELLAN VADHÅLLNINGSBESTÄMMELSER ATG (SVERIGE) OCH PHUMELELA (SYDAFRIKA)
SAMMANFATTNING AV SKILLNADER MELLAN VADHÅLLNINGSBESTÄMMELSER ATG (SVERIGE) OCH PHUMELELA (SYDAFRIKA) Spelet till sydafrikansk galopplopp sker i gemensam pool med Phumelela i Sydafrika. Det är således de
Läs merPRESS FÄLLKONSTRUKTION FOLDING INSTRUCTIONS
PRESS FÄLLKONSTRUKTION FOLDING INSTRUCTIONS Vänd bordet upp och ner eller ställ det på långsidan. Tryck ner vid PRESS och fäll benen samtidigt. Om benen sitter i spänn tryck benen mot kortsidan före de
Läs merMönster. Ulf Cederling Växjö University Ulf.Cederling@msi.vxu.se http://www.msi.vxu.se/~ulfce. Slide 1
Mönster Ulf Cederling Växjö University UlfCederling@msivxuse http://wwwmsivxuse/~ulfce Slide 1 Beskrivningsmall Beskrivningsmallen är inspirerad av den som användes på AG Communication Systems (AGCS) Linda
Läs merBridging the gap - state-of-the-art testing research, Explanea, and why you should care
Bridging the gap - state-of-the-art testing research, Explanea, and why you should care Robert Feldt Blekinge Institute of Technology & Chalmers All animations have been excluded in this pdf version! onsdag
Läs merTentamen i Matematik 2: M0030M.
Tentamen i Matematik 2: M0030M. Datum: 203-0-5 Skrivtid: 09:00 4:00 Antal uppgifter: 2 ( 30 poäng ). Examinator: Norbert Euler Tel: 0920-492878 Tillåtna hjälpmedel: Inga Betygsgränser: 4p 9p = 3; 20p 24p
Läs merSalmonella control in pig production in Sweden. Helene Wahlström, Zoonosiscenter, SVA
Salmonella control in pig production in Sweden Helene Wahlström, Zoonosiscenter, SVA s ZoonosCenter Historik salmonella Imp. Alvesta 9000 90 1:st 1:st salmonellaregulation SWEDISH SALMONELLA CONTROL PROGRAMMES
Läs merDatasäkerhet och integritet
Chapter 4 module A Networking Concepts OSI-modellen TCP/IP This module is a refresher on networking concepts, which are important in information security A Simple Home Network 2 Unshielded Twisted Pair
Läs merSchenker Privpak AB Telefon VAT Nr. SE Schenker ABs ansvarsbestämmelser, identiska med Box 905 Faxnr Säte: Borås
Schenker Privpak AB Interface documentation for web service packageservices.asmx 2012-09-01 Version: 1.0.0 Doc. no.: I04304b Sida 2 av 7 Revision history Datum Version Sign. Kommentar 2012-09-01 1.0.0
Läs merPharmacovigilance lagstiftning - PSUR
Pharmacovigilance lagstiftning - PSUR Karl Mikael Kälkner Tf enhetschef ES1 EUROPAPARLAMENTETS OCH RÅDETS DIREKTIV 2010/84/EU av den 15 december om ändring, när det gäller säkerhetsövervakning av läkemedel,
Läs merIntroduktion ICAO-EASA.
Introduktion ICAO-EASA. SSP= State Safety Program ( krav på stater från ICAO) talar bl.a. om SPI. 1 Info om kommande SMS-krav för POA. Sverige har som medlemsland i ICAO åtagit sig att ta fram ett nationellt
Läs merMOLECULAR SHAPES MOLECULAR SHAPES
Molecules with 2 electron pair groups around Linear molecules have polar bonds, but are the central atom form a linear shape. usually non-polar. is 180 linear 2 electron pairs around the central atom 1
Läs merOm oss DET PERFEKTA KOMPLEMENTET THE PERFECT COMPLETION 04 EN BINZ ÄR PRECIS SÅ BRA SOM DU FÖRVÄNTAR DIG A BINZ IS JUST AS GOOD AS YOU THINK 05
Om oss Vi på Binz är glada att du är intresserad av vårt support-system för begravningsbilar. Sedan mer än 75 år tillverkar vi specialfordon i Lorch för de flesta olika användningsändamål, och detta enligt
Läs merIs it possible to protect prosthetic reconstructions in patients with a prefabricated intraoral appliance?
r Is it possible to protect prosthetic reconstructions in patients with a prefabricated intraoral appliance? - A pilot study Susan Sarwari and Mohammed Fazil Supervisors: Camilla Ahlgren Department of
Läs merBoiler with heatpump / Värmepumpsberedare
Boiler with heatpump / Värmepumpsberedare QUICK START GUIDE / SNABBSTART GUIDE More information and instruction videos on our homepage www.indol.se Mer information och instruktionsvideos på vår hemsida
Läs merBBT057/ BBC057 BBCD057/ BBT057-NL HOLDEN COLORADO 9/2016+ HOLDEN TRAILBLAZER WD & 4WD Models
INSTALLATION GUIDE BBT057/ BBC057 BBCD057/ BBT057-NL HOLDEN COLORADO 9/2016+ HOLDEN TRAILBLAZER 2017+ 2WD & 4WD Models Ironman 4x4 BBT/ BBC/ BBCD/BBT057-NL Bull Bars fit to a Holden Colorado 9/2016+ It
Läs merTime (min)
3 2.6 TEF30 VIPP1 Fold change 2.2 1.8 1.4 1 0.6-60 0 60 120 180 240 300 360 420 480 Time (min) Supplemental Figure S1. TEF30 protein accumulates in Chlamydomonas cells exposed to high light. Chlamydomonas
Läs merBOENDEFORMENS BETYDELSE FÖR ASYLSÖKANDES INTEGRATION Lina Sandström
BOENDEFORMENS BETYDELSE FÖR ASYLSÖKANDES INTEGRATION Lina Sandström Frågeställningar Kan asylprocessen förstås som en integrationsprocess? Hur fungerar i sådana fall denna process? Skiljer sig asylprocessen
Läs merSUPPLEMENTARY INFORMATION
SUPPLEMENTARY INFORMATION SUPPLEMENTARY METHODS Preparation of the cells for transmission electron microscopy - Cells grown on coverslips were fixed for 45 minutes with 2.5% glutaraldehyde (50 mm cacodylate
Läs mer1. Varje bevissteg ska motiveras formellt (informella bevis ger 0 poang)
Tentamen i Programmeringsteori Institutionen for datorteknik Uppsala universitet 1996{08{14 Larare: Parosh A. A., M. Kindahl Plats: Polacksbacken Skrivtid: 9 15 Hjalpmedel: Inga Anvisningar: 1. Varje bevissteg
Läs merFORTA M315. Installation. 218 mm.
1 Installation 2 1 2 1 218 mm. 1 2 4 5 6 7 8 9 2 G, G0= Max 100 m 1.5 mm² (AWG 15) X1, MX, Y, VH, VC = Max 200 m 0.5 mm² (AWG 20) Y X1 MX VH VC G1 G0 G 0 V 24 V~ IN 0-10 0-5, 2-6 60 s OP O 1 2 4 5 6 7
Läs merKurskod: TAMS11 Provkod: TENB 28 August 2014, 08:00-12:00. English Version
Kurskod: TAMS11 Provkod: TENB 28 August 2014, 08:00-12:00 Examinator/Examiner: Xiangfeng Yang (Tel: 070 2234765) a. You are permitted to bring: a calculator; formel -och tabellsamling i matematisk statistik
Läs merAdding active and blended learning to an introductory mechanics course
Adding active and blended learning to an introductory mechanics course Ulf Gran Chalmers, Physics Background Mechanics 1 for Engineering Physics and Engineering Mathematics (SP2/3, 7.5 hp) 200+ students
Läs merINDUKTIV SLINGDETEKTOR INDUCTIVE LOOP DETECTOR
INDUKTIV SLINGDETEKTOR INDUCTIVE LOOP DETECTOR Slingdetektorn används som ett alternativ till mekaniska gränslägen, momentbrytare eller annat gränsläge i gödselrännor. Detektorn är kopplad till en trådslinga
Läs merUtfärdad av Compiled by Tjst Dept. Telefon Telephone Datum Date Utg nr Edition No. Dokumentnummer Document No.
Utfärdad av Compiled by Tjst Dept. Telefon Telephone David Andersson BUM 733 684 Stämpel/Etikett Security stamp/label ÅTDRAGNINGSMOMENT TIGHTENING TORQUE Granskad av Reviewed by Göran Magnusson Tjst Dept.
Läs merEASA Standardiseringsrapport 2014
EASA Standardiseringsrapport 2014 Inför EASA Standardiseringsinspektion hösten 2016 Presentatör Johan Brunnberg, Flygteknisk Inspektör & Del-M Koordinator Sjö- och luftfartsavdelningen Enheten för operatörer,
Läs merASSESSMENT AND REMEDIATION FOR CHILDREN WITH SPECIAL EDUCATIONAL NEEDS:
ASSESSMENT AND REMEDIATION FOR CHILDREN WITH SPECIAL EDUCATIONAL NEEDS: THE ROLE OF WORKING MEMORY, COMPLEX EXECUTIVE FUNCTION AND METACOGNITIVE STRATEGY TRAINING Avdelningen för psykologi Mittuniversitetet
Läs mer00-1595. Fiat 500 2007» Fiat Panda / 4x4 2003» Fiat Panda 4x4 Climbing / 4x4 Cross 20033» 619-0300
00-1595 120 Fiat 500 2007» Fiat Panda / 4x4 2003» Fiat Panda 4x4 Climbing / 4x4 Cross 20033» 619-0300 rev. 2014-04-04 DC Congratulations on purchasing an ATS towbar Alexo Towbars Sweden offer quality towbars
Läs merDen framtida redovisningstillsynen
Den framtida redovisningstillsynen Lunchseminarium 6 mars 2015 Niclas Hellman Handelshögskolan i Stockholm 2015-03-06 1 Källa: Brown, P., Preiato, J., Tarca, A. (2014) Measuring country differences in
Läs merKristina Säfsten. Kristina Säfsten JTH
Att välja metod några riktlinjer Kristina Säfsten TD, Universitetslektor i produktionssystem Avdelningen för industriell organisation och produktion Tekniska högskolan i Jönköping (JTH) Det finns inte
Läs mer