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1.
The nucleotide sequence of serine dehydratase mRNA of rat liver has been determined from a recombinant cDNA clone, previously cloned in this laboratory, and from a recombinant cDNA clone screened from a primer-extended cDNA library. The sequence of 1322 nucleotides includes the entire protein coding region and noncoding regions on the 3'- and 5'-sides. The deduced polypeptide consists of 327 amino acid residues with a calculated molecular mass of 34,462 Da. Comparison of the amino acid sequences of the serine dehydratase polypeptide with those of biosynthetic threonine dehydratase of yeast and biodegradative threonine dehydratase of E. coli revealed various extents of homology. A heptapeptide sequence, Gly-Ser-Phe-Lys-Ile-Arg-Gly, which is the pyridoxal-binding site in the yeast and E. coli threonine dehydratases was found as a highly conserved sequence. 相似文献
2.
J S Okkels H V Scheller I Svendsen B L M?ller 《The Journal of biological chemistry》1991,266(11):6767-6773
Photosystem I in barley contains a polypeptide with an apparent molecular mass of 14 kDa. The polypeptide is N-terminally blocked to amino acid sequencing, but partial amino acid sequences have been determined from three fragments obtained by chemical and enzymatic cleavage. Using an oligonucleotide probe specifying this amino acid sequence, a full length cDNA clone was isolated. The deduced amino acid sequence does not correspond to any previously identified photosystem I subunit. We designate the novel photosystem I subunit PSI-L and the corresponding nuclear gene PsaL. The cDNA clone encodes a precursor polypeptide of 209 amino acid residues with a deduced molecular mass of 22,210 Da. The precursor has a transit peptide typical of proteins imported into chloroplasts. Based on a putative maturation site, the deduced molecular mass of the mature protein is 18 kDa. The PSI-L polypeptide is hydrophobic and predicted to have at least two membrane-spanning alpha-helices. Northern blot analysis shows that the expression of the PsaL gene is light-induced similar to other of the barley photosystem I genes. Southern blot analysis indicates that PsaL is a single copy gene. Partial amino acid sequences of an N-terminally blocked 9-kDa polypeptide show high sequence similarity to the PSI-G polypeptide of spinach and Chlamydomonas reinhardtii. The gene product of PsaG in spinach has previously been assigned as subunit V (Steppuhn, J., Hermans, J., Nechushtai, R., Ljungberg, U., Thümmler, F., Lottspeich, F., and Herrmann, R. G. (1988) FEBS Lett. 237, 218-224). The present study suggests that PSI-L is equivalent to subunit V and that PSI-G is a subunit migrating closely to PSI-H (subunit VI) and PSI-C (subunit VII). 相似文献
3.
A cDNA clone encoding a 15.501 Da photosystem I (PSI) subunit of barley was isolated using an oligonucleotide based on the NH2-terminal amino acid sequence of the isolated protein. The polypeptide, which migrates with an apparent molecular mass of 9.5 kDa on denaturing SDS-PAGE, has been designated PSI-N, and the corresponding gene is PsaN. Analysis of the deduced protein sequence indicates a mature protein of 85 amino acid residues and a molecular mass of 9818 Da. PSI-N is a hydrophilic, extrinsic protein with no predicted membrane-spanning regions. The transit peptide of 60 residues (5683 Da) contains a predicted hydrophobic -helix, suggesting that the protein is routed into the thylakoid lumen. Thus, PSI-N is the second known lumenal protein component associated with PSI, together with PSI-F. 相似文献
4.
Two cDNA clones for the barley photosystem I polypeptide which migrates with an apparent molecular mass of 9.5 kDa on SDS-polyacrylamide gels have been isolated using antibodies and an oligonucleotide probe. The determined N-terminal amino acid sequence for the mature polypeptide confirms the identification of the clones. The 644 base-pair sequence of one of the clones contains one large open reading frame coding for a 14 882 Da precursor polypeptide. The molecular mass of the mature polypeptide is 10 193 Da. The hydropathy plot of the polypeptide shows one membrane-spanning region with a predicted -helix secondary structure. The gene for the 9.5 kDa polypeptide has been designated PsaH. 相似文献
5.
Primary structure, in vitro expression and import into mitochondria of a 29/21-kDa subunit of complex I from Neurospora crassa 总被引:1,自引:0,他引:1
A Videira M Tropschug S Werner 《Biochemical and biophysical research communications》1990,166(1):280-285
A full-length cDNA clone coding for a cytoplasmically-synthesized subunit of complex I from Neurospora crassa (apparent molecular mass of 29 kDa) was isolated. DNA sequencing revealed an open reading frame coding for a protein containing 201 amino acids. A molecular mass of 21323 Da was calculated. The precursor polypeptide was efficiently expressed in vitro and imported into isolated mitochondria. It is synthesized without a cleavable signal sequence and needs a membrane potential in order to bind to the mitochondrial membranes. 相似文献
6.
E Wachter K Hochstrasser 《Hoppe-Seyler's Zeitschrift für physiologische Chemie》1981,362(10):1351-1355
An acid-resistant trypsin inhibitor from human urine and serum is released in vivo by limited proteolysis from the high molecular acid-labile inter-alpha-trypsin inhibitor. The inhibitor shows an apparent molecular mass of 30 000 Da and is composed of two Kunitz-type domains. The domains are released in vitro by prolonged tryptic hydrolysis. The C-terminal domain is responsible for antitryptic activity. For the other domain no inhibitory activity towards proteinases, i.e. chymotrypsin, trypsin, pancreatic and leucocytic elastase has been demonstrated so far. The polypeptide chain comprising both domains consists of 122 residues and has a molecular mass of only 13 400 Da. In this work we have found that both, the N-terminal extension peptide with 21 residues and the "inactive" domain are linked O-glycosidically and N-glycosidically, respectively, with large carbohydrate moieties. The N-terminal amino acid sequence of the human urinary trypsin inhibitor was determined by solid-phase Edman degradation of a single peptide. The molecular mass calculated for the total polypeptide chain of 143 residues should be 15 340 Da; from the difference to the measured value (30 000 Da) it is concluded that the glycopeptide contains a considerable carbohydrate moiety. 相似文献
7.
Molecular cloning and nucleotide sequence of cDNA encoding the human liver S-adenosylmethionine synthetase. 总被引:4,自引:0,他引:4
cDNA clone for human liver S-adenosylmethionine synthetase (liver-specific isoenzyme) was isolated from a cDNA library of human liver poly(A)+ RNA. The cDNA sequence encoded a polypeptide consisting of 395 amino acid residues with a calculated molecular mass of 43675 Da. Alignment of the predicted amino acid sequence of this protein with that of rat liver S-adenosylmethionine synthetase showed a high degree of similarity. The coding region of the human liver S-adenosylmethionine synthetase cDNA sequence was 89% identical at the nucleotide level and 95% identical at the amino acid level to the sequence for rat liver S-adenosylmethionine synthetase. 相似文献
8.
The key regulatory enzyme in the biosynthetic pathway of the plant hormone ethylene is 1-aminocyclopropane-1-carboxylic acid (ACC) synthase (EC 4.4.1.14). We have partially purified ACC synthase 6,000-fold from Cucurbita fruit tissue treated with indoleacetic acid + benzyladenine + aminooxyacetic acid + LiCl. The enzyme has a specific activity of 35,000 nmol/h/mg protein, a pH optimum of 9.5, an isoelectric point of 5.0, a Km of 17 microM with respect to S-adenosylmethionine, and is a dimer of two identical subunits of approximately 46,000 Da each. The subunit exists in vivo as a 55,000-Da species similar in size to the primary in vitro translation product. DNA sequence analysis of the cDNA clone pACC1 revealed that the coding region of the ACC synthase mRNA spans 493 amino acids corresponding to a 55,779-Da polypeptide; and expression of the coding sequence (pACC1) in Escherichia coli as a COOH terminus hybrid of beta-galactosidase or as a nonhybrid polypeptide catalyzed the conversion of S-adenosylmethionine to ACC (Sato, T., and Theologis, A. (1989) Proc. Natl. Acad. Sci. U.S.A. 86, 6621-6625). Immunoblotting experiments herein show that the molecular mass of the beta-galactosidase hybrid polypeptide is 170,000 Da, and the size of the largest nonhybrid polypeptide is 53,000 Da. The data suggest that the enzyme is post-translationally processed during protein purification. 相似文献
9.
cDNA cloning of porcine brain prolyl endopeptidase and identification of the active-site seryl residue 总被引:8,自引:0,他引:8
Prolyl endopeptidase is a cytoplasmic serine protease. The enzyme was purified from porcine kidney, and oligonucleotides based on peptide sequences from this protein were used to isolate a cDNA clone from a porcine brain library. This clone contained the complete coding sequence of prolyl endopeptidase and encoded a polypeptide with a molecular mass of 80,751 Da. The deduced amino acid sequence of prolyl endopeptidase showed no sequence homology with other known serine proteases. [3H]Diisopropyl fluorophosphate was used to identify the active-site serine of prolyl endopeptidase. One labeled peptide was isolated and sequenced. The sequence surrounding the active-site serine was Asn-Gly-Gly-Ser-Asn-Gly-Gly. This sequence is different from the active-site sequences of other known serine proteases. This difference and the lack of overall homology with the known families of serine proteases suggest that prolyl endopeptidase represents a new type of serine protease. 相似文献
10.
Neuropeptide Y (NPY) has been isolated from brain extracts of the rainbow trout (Oncorhynchus mykiss) and subjected to structural analyses. Plasma desorption mass spectroscopy estimated the molecular mass of the purified peptide as 4303.9 Da. Automated Edman degradation unequivocally established the sequence of a 36 amino acid residue peptide as: Tyr-Pro-Pro-Lys-Pro-Glu-Asn-Pro-Gly-Glu-Asp-Ala-Pro-Pro-Glu-Glu-Leu-Ala-Lys-Tyr-Tyr-Ala-Leu-Arg-His-Tyr-Ile-Asn-Leu-Ile-Thr-Arg-Gln-Arg-Tyr. The molecular mass calculated from this sequence (4304 Da) is consistent with that obtained by mass spectroscopy. The presence of a C-terminal amide was established by radioimmunoassay. Rainbow trout NPY is identical in primary structure to coho salmon (Oncorhynchus kisutch) pancreatic polypeptide (PP). These data may indicate that, in this group of salmonid fishes, a single member of the NPY/PP peptide family is expressed in both neurons and peripheral endocrine cells. 相似文献
11.
12.
Cavada BS da Silva LI Ramos MV Galvani FR Grangeiro TB Leite KB Assreuy AM Cajazeiras JB Calvete JJ 《Protein and peptide letters》2003,10(6):607-617
A glucose/mannose lectin was purified by affinity chromatography from Pisum arvense seeds (PAL) and the 50 kDa molecular mass in solution determined by size exclusion chromatography. SDS-PAGE and electrospray ionization mass spectrometry showed two distinct polypeptide chains: alpha (Mr. 5591 Da) and beta (19986 Da). The lectin was extensively characterized in terms of its biochemical and biological aspects. The amino acid sequence was established by Edman degradation of overlapping peptides. PAL in solution behaves as a dimer and has its monomeric structure formed by two distinct polypeptide chains named alpha (Mr. 5591 Da) and beta (19986 Da) by Electrospray ionization (ESI) mass spectrometry. PAL possesses identical amino acid sequences to that of pea seed lectin but undoubtedly does not exhibit sequence heterogeneity. It is discussed that P. arvense should be considered as a synonym of P. sativum. Furthermore, like pea lectin, PAL discriminates biantennary fucosylated glycan, determined by surface plasmon resonance. 相似文献
13.
The primary structure of a 4.0-kDa photosystem I polypeptide encoded by the chloroplast psaI gene 总被引:2,自引:0,他引:2
H V Scheller J S Okkels P B H?j I Svendsen P Roepstorff B L M?ller 《The Journal of biological chemistry》1989,264(31):18402-18406
Partial amino acid sequences have been determined for a 4.0-kDa photosystem I polypeptide from barley. A comparison with the sequence of the chloroplast genome of Nicotiana tabacum and Marchantia polymorpha identified the polypeptide as chloroplast-encoded. We designate the corresponding gene psaI and the polypeptide PSI-I. The barley chloroplast psaI gene was sequenced. The gene encodes a polypeptide of 36 amino acid residues with a deduced molecular mass of 4008 Da. The 4.0-kDa polypeptide is N-terminally blocked with a formyl-methionine residue. Plasma desorption mass spectrometry established that the polypeptide is not post-translationally processed except for possible conversion of a methionine residue into methionine sulfone. The hydrophobic 4.0-kDa polypeptide is predicted to have one membrane-spanning alpha-helix and is homologous to transmembrane helix E of the D2 reaction center polypeptide of photosystem II. 相似文献
14.
Isolation, sequence, and expression in Escherichia coli of the Pseudomonas sp. strain ACP gene encoding 1-aminocyclopropane-1-carboxylate deaminase. 总被引:1,自引:0,他引:1 下载免费PDF全文
R E Sheehy M Honma M Yamada T Sasaki B Martineau W R Hiatt 《Journal of bacteriology》1991,173(17):5260-5265
Pseudomonas sp. strain ACP is capable of growth on 1-aminocyclopropane-1-carboxylate (ACC) as a nitrogen source owing to induction of the enzyme ACC deaminase and the subsequent conversion of ACC to alpha-ketobutyrate and ammonia (M. Honma, Agric. Biol. Chem. 49:567-571, 1985). The complete amino acid sequence of purified ACC deaminase was determined, and the sequence information was used to clone the ACC deaminase gene from a 6-kb EcoRI fragment of Pseudomonas sp. strain ACP DNA. DNA sequence analysis of an EcoRI-PstI subclone demonstrated an open reading frame (ORF) encoding a polypeptide with a deduced amino acid sequence identical to the protein sequence determined chemically and a predicted molecular mass of 36,674 Da. The ORF also contained an additional 72 bp of upstream sequence not predicted by the amino acid sequence. Escherichia coli minicells containing the 6-kb clone expressed a major polypeptide of the size expected for ACC deaminase which was reactive with ACC deaminase antiserum. Furthermore, a lacZ fusion with the ACC deaminase ORF resulted in the expression of active enzyme in E. coli. ACC is a key intermediate in the biosynthesis of ethylene in plants, and the use of the ACC deaminase gene to manipulate this pathway is discussed. 相似文献
15.
A cysteine-rich polypeptide, termed CRP1, with a molecular mass of 5829 Da was found to occur in the mid-gut gland of the scallop Patinopecten yessoensis. CRP1 was purified by reverse phase and cation-exchange chromatographies. The amino acid sequence of CRP1 was deduced from its N-terminal amino acid sequence, amino acid composition and the sequence of a partial cDNA, indicating that CRP1 is a 57-amino-acid polypeptide containing 12 cysteine residues with a calculated molecular mass of 5841 Da (5829 Da when oxidized to form six disulfide bridges). A homology search of databases revealed that the deduced amino acid sequence of CRP1 displays significant similarity to those of granulin/epithelins, a family of growth-modulating factors; all cysteine residues in CRP1 are located at the same positions as those conserved characteristically in other known granulin/epithelins. Purified CRP1 inhibited the proliferation of mouse embryo cells. The results suggest that CRP1 functions as a growth-modulating factor in the scallop, and that granulin/epithelin family polypeptides and their precursors play physiologically important roles in invertebrates. 相似文献
16.
Complete amino acid sequence of endo-beta-N-acetylglucosaminidase from Flavobacterium sp 总被引:2,自引:0,他引:2
K Takegawa B Mikami S Iwahara Y Morita K Yamamoto T Tochikura 《European journal of biochemistry》1991,202(1):175-180
The complete amino acid sequence of endo-beta-N-acetylglucosaminidase from Flavobacterium sp. has been determined by analysis of peptides after cleavage with lysyl endopeptidase, pepsin and chymotrypsin. The protein consists of a single polypeptide chain consisting of 267 amino acid residues and a molecular mass of 27972 Da. The sequence of Flavobacterium endo-beta-N-acetylglucosaminidase is very close to that of the Streptomyces enzyme (endo-H), having 60% similarity and very similar hydropathy profiles. Similarities were also found between Flavobacterium endo-beta-N-acetylglucosaminidase and chitinases from Bacillus circulans, Serratia marcescens and Phaseolus vulgaris. 相似文献
17.
Chloroplast NADH dehydrogenase from Pisum sativum: characterization of its activity and cloning of ndhK gene 总被引:1,自引:0,他引:1
The pea chloroplast ndhK gene coding for a component of a NADH-plastoquinone oxidoreductase has been cloned and sequenced. This gene codes for a polypeptide of 227 amino acids and a predicted molecular mass of 25,495 Da which belongs to the family of the 20 kDa PSST subunit of the bovine mitochondrial complex I. A fragment of this gene has been overexpressed in Escherichia coli, and antibodies against the expressed polypeptide recognize a protein of the predicted molecular mass from pea thylakoid membranes. This polypeptide is a component of a protein complex with NADH dehydrogenase activity and is not associated with ferredoxin-NADP+ reductase. 相似文献
18.
Amino-acid sequence of the smallest protein of the cytochrome c1 subcomplex from beef heart mitochondria 总被引:2,自引:0,他引:2
H Sch?gger G von Jagow U Borchart W Machleidt 《Hoppe-Seyler's Zeitschrift für physiologische Chemie》1983,364(3):307-311
A cytochrome c1 subcomplex was obtained from purified complex III. The subcomplex consists of three protein subunits, present at an equimolar stoichiometry. The primary structures of two of these proteins, the heme-carrying cytochrome c1 and a protein with a molecular mass of 9175 Da, have been published by Wakabayashi et al. The covalent structure of the smallest subunit is presented in this paper. This protein consists of 62 amino acid residues; its molecular mass was calculated to be 7189 Da. The sequence was determined by complete solid-phase Edman degradation of the uncleaved polypeptide and was confirmed by sequencing the C-terminal fragment resulting from cleavage of a single tryptophyl bond. 相似文献
19.
Molecular cloning and characterization of nlpH, encoding a novel, surface-exposed, polymorphic, plasmid-encoded 33-kilodalton lipoprotein of Borrelia afzelii. 下载免费PDF全文
M Theisen 《Journal of bacteriology》1996,178(22):6435-6442
Borrelia burgdorferi sensu lato organisms, comprising B. burgdorferi sensu stricto, Borrelia afzelii, and Borrelia garinii, are tick-borne pathogens causing Lyme borreliosis in humans. To identify putative virulence determinants, a B. afzelii DNA library was screened for Congo red dye binding, a property associated with virulence in pathogenic bacteria. One clone was found to carry a 663-nucleotide-long open reading frame encoding a Congo red dye-binding protein with a calculated molecular mass of 25,660 Da. The amino acid sequence deduced from its nucleotide sequence was found to include a consensus bacterial lipidation site present at residues 15 to 18 (Leu-Ser-Gly-Cys). The lipoprotein nature was demonstrated by incorporation of radioactive palmitate; hence, this protein has been termed NlpH, for new lipoprotein H. NlpH is located on the surface of B. afzelii, and the nlpH gene is found on a circular plasmid. The nlpH gene is also found in B. burgdorferi sensu stricto and B. garinii. Immediately upstream of nlpH is located a smaller reading frame encoding a polypeptide containing the casein kinase II phosphorylation recognition sequence, (Ser/Thr)-X-Y-(Glu/Asp), repeated 10 times. 相似文献