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1.
GILLES MICHEL BRIGITTE LVY MARIE-THRSE CHAUVET JACQUELINE CHAUVET ROGER ACHER 《Chemical biology & drug design》1990,36(3):302-307
Vasotocin-associated neurophysin (MSEL-neurophysin) has been purified from goose neurohypophysis through molecular sieving and high-pressure reverse-phase liquid chromatography (HPLC). The protein has a molecular mass (measured by SDS-polyacrylamide gel electrophoresis) of 17kDa in contrast to 10kDa found for the mammalian MSEL-neurophysins. Complete amino acid sequence (131 residues) has been determined mainly through tryptic or staphylococcal proteinase peptides derived from carboxyamido-methylated neurophysin, isolated by HPLC and microsequenced. N- and C-terminal sequences have been established by Edman degradation or action of carboxypeptidase Y, respectively, applied on the native protein. Goose MSEL-neurophysin is homologous to the two-domain “big” MSEL-neurophysin previously identified in the frog. It appears that in non-mammalian tetrapods, namely birds and amphibians, the proteolytic processing of the pro-vasotocin involves only one cleavage, releasing the hormone moiety and a “big” neurophysin with two domains homologous to mammalian MSEL-neurophysin and copeptin, respectively. Comparison of the avian protein with its mammalian and amphibian counterparts reveals that the first half of the polypeptide chain is evolutionarily much less variable than the second and that the goose protein resembles the frog protein much more than the mammalian one. 相似文献
2.
C. LAZURE H.S. SAAYMAN R.J. NAUD W. OELOFSEN M. CHRTIEN 《Chemical biology & drug design》1987,30(5):634-645
The neurohypophyseal hormones vasopressin and oxytocin are known to be synthesized in eutherian mammals as part of larger precursors containing either MSEL-or VLDV-neurophysins. A neurophysin has been isolated from ostrich neurohypophyses and shown by partial amino acid sequence determination to be related to mammalian VLDV-neurophysin. The present report describes the complete amino acid sequence of this ostrich neurophysin containing 93 residues. This amino acid sequence, the first reported in birds, differs in a remarkable manner from its mammalian homolog. Indeed, it contains a large number of substitutions, including one insertion, distributed throughout the polypeptide chain when compared to known VLDV-neurophysins. Whereas many of these substitutions are localized inside the so-called constant region of the neurophysin, the highest variation can be found in the COOH-terminal region. 相似文献
3.
The complete amino acid sequence of growth hormone from fin whale pituitary has been determined. It consists of 190 amino acid residues with two disulfide bridges formed by residues 52–163 and 180–188. The sequence identity with sei whale, bovine, ovine and human growth hormone is 90%, 91%, 90% and 68%, respectively. 相似文献
4.
5.
C. LAZURE H.S. SAAYMAN R.J. NAUD W. OELOFSEN M. CHRTIEN 《Chemical biology & drug design》1989,33(1):46-58
Mammalian neurohypophyseal hormones, oxytocin and vasopressin, are known to be synthesized as part of two larger precursors containing, respectively, a VLDV-neurophysin and a MSEL-neurophysin together with its associated glycopeptide. Starting from ostrich neurohypophyses, a “big” neurophysin was isolated and chemically characterized. Following sequence determination of the CNBr-derived fragments and of peptides obtained from trypsin and V8-protease digestion of the oxidized protein, this “big” neurophysin was found to contain an MSEL-neurophysin moiety (94 residues) still covalently associated with the COOH-terminal glycopeptide (38 residues, copeptin). This study demonstrates that the ostrich MSEL-neurophysin sequence closely resembles all known MSEL-neurophysin sequences and that, furthermore, it does not contain the single amino acid insertion shown previously in the ostrich VLDV-neurophysin. It is also shown that the stretch of amino acids, linking the MSEL-neurophysin and the copeptin, is clearly different from its mammalian homologues and lacks the Arg residue normally recognized by the cleaving enzyme. This study also demonstrates that the ostrich copeptin is more closely related to the amphibian copeptin sequence than to its mammalian homologue, leading to the hypothesis that two families of copeptin molecules might exist. Thus, the ostrich MSEL-neurophysin-copeptin molecule is the first “big” neurophysin reported in birds and, together with the guinea pig and amphibian homologues, represents the third example of partial or no neurophysin-copeptin cleavage. 相似文献
6.
A subunit of molecular weight 21 000 from arachin, the major peanut protein, was isolated in pure form and primary structure was determined. The subunit was fragmented with CNBr, trypsin, and NBS; the fragments were separated and isolated by PAGE, gel filtration, Dowex treatment, and paper electrophoresis, and Edman degradation on each fragment, including the intact subunit, was performed. The PTH-amino acids thus obtained were identified by UV spectroscopy and TLC. The complete sequence of 176 residues was established by overlapping technique. 相似文献
7.
A subunit (M, 15 600) from the high molecular weight protein from rapeseed was separated and isolated; its purity and homogeneity were ascertained. The subunit was cleaved with cyanogen bromide, trypsin, chymotrypsin, and Staphylococcus aureus V8 protease. The fragments were separated and isolated by polyacrylamide gel electrophoresis, gel filtration, column chromatography on Dowex 1 × 2, and paper electrophoresis. The amino acid compositions of the intact subunit and different fragments obtained from enzymatic and chemical cleavages were determined. The subunit and its fragments were sequenced by manual Edman method. The phenylthiohydantoin amino acids obtained after each step were identified by thin-layer chromatography and ultraviolet spectroscopy. The complete amino acid sequence of the subunit consisting of 125 amino acid residues has been established by the overlapping method. 相似文献
8.
The partial amino acid sequence of porcine elastase II, isolated from crude trypsin Type II, was determined. The enzyme consists of two polypeptide chains, a light chain composed of 11 residues, and a heavy chain (Mr = 23 500) with four intrachain disulfide bridges; the two chains are held together by one interchain disulfide bond. Elastase II was fragmented into several peptides by chemical cleavages with CNBr at the two methionine residues, 99 and 180 (chymotrypsinogen numbering), and with hydroxylamine at the peptide bond following DIP-Ser195. About 50% of the sequence was determined and the positions of 120 amino acids were located, including the light chain residues and most of the active site residues. The partial sequence shows 64% difference between porcine elastase II and elastase I and only 26% difference between porcine elastase II and bovine chymotrypsin B. 相似文献
9.
Andre Silvanovich Margaret A Nemeth Ping Song Rod Herman Laura Tagliani Gary A Bannon 《Toxicological sciences》2006,90(1):252-258
Typically, genetically engineered crops contain traits encoded by one or a few newly expressed proteins. The allergenicity assessment of newly expressed proteins is an important component in the safety evaluation of genetically engineered plants. One aspect of this assessment involves sequence searches that compare the amino acid sequence of the protein to all known allergens. Analyses are performed to determine the potential for immunologically based cross-reactivity where IgE directed against a known allergen could bind to the protein and elicit a clinical reaction in sensitized individuals. Bioinformatic searches are designed to detect global sequence similarity and short contiguous amino acid sequence identity. It has been suggested that potential allergen cross-reactivity may be predicted by identifying matches as short as six to eight contiguous amino acids between the protein of interest and a known allergen. A series of analyses were performed, and match probabilities were calculated for different size peptides to determine if there was a scientifically justified search window size that identified allergen sequence characteristics. Four probability modeling methods were tested: (1) a mock protein and a mock allergen database, (2) a mock protein and genuine allergen database, (3) a genuine allergen and genuine protein database, and (4) a genuine allergen and genuine protein database combined with a correction for repeating peptides. These analyses indicated that searches for short amino acid sequence matches of eight amino acids or fewer to identify proteins as potential cross-reactive allergens is a product of chance and adds little value to allergy assessments for newly expressed proteins. 相似文献
10.
目的对结肠黑变病相关基因金属泛激蛋白-1(MPS-1)cDNA序列和氨基酸序列进行综合分析,为其结构和功能的研究奠定理论基础。方法从Pubmed数据库中获得目标cDNA序列,利用基因和蛋白质分析处理软件DNAMAN、NCBI ORF finder、BLAST、Conserved Domains、GOR、SWISS-MODEL等软件对该目标的基因序列和氨基酸序列进行综合分析。结果 MPS-1蛋白cDNA序列长为361 bp,编码84个氨基酸残基,基因序列比对显示该蛋白与核糖体蛋白S27(RPS27)mRNA的编码序列同源度最高,两者核苷酸的相似度为361/361(100%),氨基酸序列比对显示该蛋白与RPS27的氨基酸序列同源度最高,两者氨基酸的相似度为84/84(100%),目标序列中存在1段Ribosomal_S27e家族保守结构域,二级结构和三级结构预测显示目标蛋白中主要存在α-螺旋、β-折叠和无规卷曲的结构。结论 MPS-1蛋白与RPS27同源度高,与肿瘤的发生密切相关。 相似文献
11.
运用液质联用、两种串联质谱对融合蛋白FP3的氨基酸全序列测定, 确证其一级结构。将样品还原烷基化后, 通过胰蛋白酶酶解蛋白, PNGase F去除多肽混合物中糖肽的糖基化, 将去糖后的总肽用于液质联用系统, 通过液相分离后, 运用Q-TOF和线性离子阱两种串联质谱测定各个肽段的b, y碎片离子, 分析测定融合蛋白FP3的氨基酸全序列。通过LC-ESI-Q-TOF完成了融合蛋白FP3的76%氨基酸序列测定, 通过LC-ESI-Trap完成余下24%氨基酸序列测定。液质联用、串联质谱法测定蛋白质氨基酸序列快速、灵敏、准确, 是对重组蛋白结构分析和确证的重要手段。 相似文献
12.
将免疫亲和层析纯化的抗乙酰胆碱酯酶单克隆抗体3F3还原并烷基化后,用凝胶过滤法分离纯化抗体重链。以SDS-PAGE检定其纯度后,将完整的3F3单克隆抗体重链做氨基酸顺序分析,测定了可变区N端的42个氨基酸的顺序。 相似文献
13.
The sequence of a long-chain neurotoxin (71 amino acid residues, 10 half-cystines) from the venom of the African banded water cobra (Boulengerina annulata) was determined by Edman degradation. It exhibits high sequence similarity with long-chain neurotoxins from the venoms of four species of African cobras (genus Naja), which are collectively more similar to the Boulengerina toxin than to those of Asian Naja species. These results are discussed in the light of phylogenetic hypotheses on the relationships of African cobras. 相似文献
14.
目的通过对鲍曼不动杆菌的耐药基因和插入序列遗传标记进行研究来了解鲍曼不动杆菌的耐药机制,为临床治疗性用药、提供临床疗效提供实验依据。方法通过细菌药物敏感试验检测鲍曼不动杆菌对抗菌药物的耐药情况,通过PCR方法检测鲍曼不动杆菌的携带β-内酰胺类药物耐药基因的状况以及插入序列遗传标记的情况。结果细菌药物敏感试验检测结果是鲍曼不动杆菌对多种抗菌药物均耐药,共检出TEM、OXA-2、ADC、CTX—M-1、CTX—M-2、IMP、OXA-24等耐药基因,插入序列遗传标记ISabal、IS26、tnp513检出率分别为80.43%和47.82%、52.17%。结论β-内酰胺酶基因广泛存在于鲍曼不动杆菌中,是鲍曼不动杆菌对目前临床常用的β-内酰胺类药物存在多重耐药的主要原因。多重耐药鲍曼不动杆菌的耐药性与插人序列遗传标记有一定的相关性。 相似文献
15.
Elena V Klyshko Marina P Issaeva Margarita M Monastyrnaya Anna P Il'yna Konstantin V Guzev Tatyana I Vakorina Pavel S Dmitrenok Tatyana A Zykova Emma P Kozlovskaya 《Toxicon》2004,44(3):315-324
A new cytolytic toxin, actinoporin RTX-S II, was isolated from the sea anemone Radianthus macrodactylus with a high degree of purity by a combination of gel filtration, ion-exchange and reverse-phase chromatography. RTX-S II has molecular mass of 19,280 Da and isoelectric point of 10.0. The hemolytic activity of RTX-S II is inhibited by sphingomyelin. RTX-S II had an LD(50) of 70 mg/kg, and is lacking in phospholipase activity. The amino acid composition of this protein contains a high amount of basic and non-polar amino acids and no cysteine. The N-terminal sequence of RTX-S II was determined. The partial amino acid sequence (141 aa) of RTX-S II was deduced based on the cDNA sequence obtained with two oligonucleotides encoding the N-terminal portion of RTX-S II and the internal conserved cytolysin peptide by PCR. A comparison of the RTX-S II cDNA sequence and the rtx-s II gene obtained with the same PCR primers indicates that they are 100% identical at the nucleotide level. It shows that no introns are present in the corresponding region of the rtx-s II gene. Multiple alignments of RTX-S II with known sequences of actinoporins show that RTX-S II is highly homologous to magnificalysin II from Heteractis magnifica. The predicted secondary structure of RTX-S II is predominantly anti-parallel beta-structure, which is in good agreement with experimental data obtained from other sea anemones-actinoporins. 相似文献
16.
C.-S. LIU H.-L. LEU C.-S. CHANG S.-W. CHEN T.-B. LO 《Chemical biology & drug design》1989,34(4):257-261
Two phospholipases were found in the venom of Bungarus fasciatus, one in fraction III, the other in fraction X of the chromatographic separation. A neutral PLA2(III) purified from fraction III was subjected to amino acid sequencing by means of an automated sequenator applied to the intact RCM-PLA2 (III) and the individual peptides obtained from HPLC separation of the three types of enzymatic peptides. PLA(III) was shown to consist of 118 amino acid residues with 14 half-cystines. It is 65% homologous to the basic PLA2 obtained from fraction X. 相似文献
17.
Amino acid sequence and function of a new alpha-toxin from the Amazonian scorpion Tityus cambridgei.
Anna Rosa Murgia Cesar V F Batista Gianfranco Prestipino Lourival D Possani 《Toxicon》2004,43(6):737-740
A toxic peptide earlier denominated Tc48b [Toxicon 40 (2002) 557] was purified to homogeneity and its amino acid sequence determined. It has 64 amino acid residues stabilized by four disulfide bridges with a molecular weight of 7,385.2 atomic mass units (a.m.u.). It affects Na(+)-permeability in pituitary GH3 cells in culture, in a similar fashion as those reported for alpha-scorpion toxins, contrary to most of the New World scorpion toxins that are beta-toxins. 相似文献
18.
A basic protein cytolysin previously isolated from the Caribbean sea anemone Stichodactyla helianthus was shown by CM cellulose chromatography to consist of four isotoxins possessing different N-terminal amino acid sequences. These are designated as toxins I-IV in order of increasing isoelectric point. The estimated molecular sizes (17,400-18,200) of toxins I-III were very similar; toxins I and II posses one additional amino acid at their amino terminus relative to toxin III. Under denaturing conditions, toxin IV behaved as a significantly larger (19,600) polypeptide; Edman sequencing established that it possesses a seven residue extension at the N-terminal end relative to toxin III. None of the variants contained half-cystines or reducing sugars. Toxin III contributed 83% of the total purified cytolytic (hemolytic) activity, toxin II 14%, and the relatively insoluble toxins I and IV together only contributed about 3% of the total cytolytic activity. Cytolysin III lysed Ehrlich ascitic tumour cells, but when administered intraperitoneally in nonlethal doses to mice already inoculated with this tumour, it failed to protect the mice against the tumour. Comparison of the partial amino acid sequence of equinatoxin, another sea anemone protein cytolysin, with that of Stichodactyla cytolysin III indicates they are highly homologous. Many other cytolytic proteins isolated from sea anemones share these properties with Stichodactyla cytolysins: (1) selective inhibition of hemolytic activity by preincubation with sphingomyelin, (2) a molecular size of 10,000-20,000, and (3) an isoelectric point of 9 or above. 相似文献
19.
The most challenging task in RNA interference is the design of active small interfering RNA (siRNA) sequences. Numerous strategies have been published to select siRNA. They have proved effective in some applications but have failed in many others. Nonetheless, all existing guidelines have been devised to select effective siRNAs targeting human or murine genes. They may not be appropriate to select functional sequences that target genes from other organisms like viruses. In this study, we have analyzed 62 siRNA duplexes of 19 bases targeting three genes of three morbilliviruses. In those duplexes, we have checked which features are associated with siRNA functionality. Our results suggest that the intramolecular secondary structure of the targeted mRNA contributes to siRNA efficiency. We also confirm that the presence of at least the sequence motifs U13, A or U19, as well as the absence of G13, cooperate to increase siRNA knockdown rates. Additionally, we observe that G11 is linked with siRNA efficacy. We believe that an algorithm based on these findings may help in the selection of functional siRNA sequences directed against viral genes. 相似文献
20.
F. SAMPIERI C. HABERSETZER-ROCHAT M.-F. MARTIN C. KOPEYAN H. ROCHAT 《Chemical biology & drug design》1987,29(2):231-237
The complete amino acid sequence of toxin XI of the North African scorpion Buthus occitanus tunetanus has been elucidated by automatic sequencing of the reduced and alkylated toxin and of the peptides obtained after tryptic cleavage restricted to arginyl bonds. This toxin is structurally homologous to toxin II of Androctonus australis Hector, the most active among the α-toxins, but is far less potent, both in vivo and in vitro. This work points out 12 mutations, many of which are conservative. Nevertheless, the most striking difference is the replacement of the lysine residue at position 58, known to be important in the activity of AaH toxin II, by a valine residue. Thus, it seems that the presence of a positive charge at this location facilitates the interactions between the receptor on the sodium channel and the α-type toxins. 相似文献