Purification, characterization and gene cloning of Da-36, a novel serine protease from Deinagkistrodon acutus venom
文献类型：期刊论文
| 作者 | Zheng Y1,2; Ye FP2,3; Wang J2; Liao GY1; Zhang Y3; Fan QS[*]2; Lee WH[*]3 |
| 刊名 | TOXICON
 |
| 出版日期 | 2013 |
| 卷号 | 67期号:X页码:1-11 |
| 关键词 | Deinagkistrodon acutus Gene Serine protease Purification |
| 通讯作者 | fqs168@126.com ; leewh@mail.kiz.ac.cn |
| 合作状况 | 其它 |
| 英文摘要 | A serine protease termed Da-36 was isolated from crude venom of Deinagkistrodon acutus. The enzyme was a single chain protein with an apparent molecular weight of 36,000 on SDS-PAGE with an isoelectric point of 6.59. Da-36 could clot human plasma by cleaving the A alpha, B beta and gamma chains of fibrinogen and also exhibited arginine esterase activity. The proteolytic activity of Da-36 toward TAME was strongly inhibited by PMSF and moderately affected by benzamidine and aprotinin, indicating that it was a serine protease. Meanwhile. Da-36 showed stability with wide temperature (20-50 degrees C) and pH value ranges (pH 6-10). Divalent metal ions of Ca2+, Mg2+, and Mn2+ had no effects but Zn2+ and Cu2+ inhibited the arginine esterase activity of Da-36. Total DNA was extracted directly-from the lyophilized crude venom and the gene (5.5 kbp) coding for Da-36 had been successfully cloned. Sequence analysis revealed that the Da-36 gene contained five exons and four introns. The mature Da-36 was encoded by four separate exons. The deduced mature amino acid sequence of Da-36 was in good agreement with the determined N-terminal sequence of the purified protein and shared high homology with other serine proteases isolated from different snake venoms. Blast search using amino acid sequence of Da-36 against public database revealed that Da-36 showed a maximal identity of 90% with both Dav-X (Swiss-Prot: Q9I8W9.1) and thrombin-like protein 1 (GenBank: AAW56608.1) from the same snake species, indicating that Da-36 is a novel serine protease. |
| 收录类别 | SCI |
| 资助信息 | This work was supported by The National Natural Sci- ence Foundation of China (31071926), 2011CI139 from Yunnan Province and China Postdoctoral Science Founda- tion (20080440214, 200902656). |
| 语种 | 英语 |
| WOS记录号 | WOS:000320075300001 |
| 公开日期 | 2013-07-12 |
| 源URL | [http://159.226.149.42:8088/handle/152453/7568]  |
| 专题 | 昆明动物研究所_动物活性蛋白多肽组学 昆明动物研究所_动物模型与人类重大疾病机理重点实验室 |
| 作者单位 | 1.Institute of Medical Biology, Chinese Academy of Medical Sciences & Peking Union Medical College, Kunming 650118, China 2.Institute of Military Medical, Chengdu Military Region’s Center for Disease Control & Prevention, Kunming 650032, China 3.Key Laboratory of Animal Models and Human Disease Mechanisms of Chinese Academy of Sciences & Yunnan Province, Kunming Institute of Zoology, The Chinese Academy of Sciences, Kunming, Yunnan 650223, China |
| 推荐引用方式 GB/T 7714 | Zheng Y,Ye FP,Wang J,et al. Purification, characterization and gene cloning of Da-36, a novel serine protease from Deinagkistrodon acutus venom[J]. TOXICON,2013,67(X):1-11. |
| APA | Zheng Y.,Ye FP.,Wang J.,Liao GY.,Zhang Y.,...&Lee WH[*].(2013).Purification, characterization and gene cloning of Da-36, a novel serine protease from Deinagkistrodon acutus venom.TOXICON,67(X),1-11. |
| MLA | Zheng Y,et al."Purification, characterization and gene cloning of Da-36, a novel serine protease from Deinagkistrodon acutus venom".TOXICON 67.X(2013):1-11. |
入库方式： OAI收割
来源：昆明动物研究所
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