Molecular cloning and characterization of a complement-depleting factor from king cobra, Ophiophagus hannah
文献类型：期刊论文
| 作者 | Zeng L1,3; Sun QY2; Zhang Y1; Jin Y1; Zhang Y[*]1; Lee WH1 |
| 刊名 | TOXICON
 |
| 出版日期 | 2012 |
| 卷号 | 60期号:3页码:290-301 |
| 关键词 | Cobra venom factor Ophiophagus hannah Purification Molecular cloning Complement-depleting activity |
| 通讯作者 | zhangy@mail.kiz.ac.cn |
| 合作状况 | 其它 |
| 英文摘要 | Cobra venom factor (CVF) is an anti-complement factor existing in cobra venom. CVF proteins have been purified from the venoms of Naja haje, Naja siamensis, Naja atra, Naja kaouthia, Naja naja, Naja melanoleuca and Austrelaps superbus, but only three full-length cDNA sequences of CVF are available. In the present work, a cobra venom factor termed OVF was purified from the crude venom of Ophiophagus hannah by successive gel filtration, ion-exchange and heparin affinity chromatography steps. The purified OVF was homogenous on the SDS-PAGE gel with an apparent molecular weight of 140 kDa under non-reducing conditions. Under reducing conditions, OVF was divided into three bands with apparent molecular weight of 72 kDa (alpha chain), 45 kDa (beta chain) and 32 kDa (gamma chain), respectively. OVF consumed complement components with anti-complement activity of 154 units per mg. By using Reverse transcription-PCR and 5'-RACE assay, the open reading frame of OVF was obtained. MALDI-TOF and protein sequencing assays confirmed the cloned cDNA coding for OVF protein. The cDNA sequence of OVF is conservative when aligned with that of other CVFs. Phylogenetic analysis revealed OVF is closer to CVF from N. kaouthia than to AVF-1 and AVF-2 from A. superbus. Our results demonstrated that OVF has its unique features as following: 1) The N-terminal amino acid sequence of OW gamma chain is different from that of other known CVFs, suggesting that the OVF gamma chain might be further processed; 2) Unlike N. kaouthia CVF and A. superbus AVF-1, which have potential N-linked glycosylation sites located in both alpha and beta chain, OW only has N-linked glycosylation site in its alpha chain as revealed by Schiff's reagent staining and protein sequence analysis; 3) In addition to the 27 well conserved cysteine residues in all known CVFs. OVF have an additional cysteine residue in its gamma chain. Understanding the importance of above mentioned specific characteristics might provide useful information on structure-function relationship between CVF and complement system. |
| 资助信息 | This work was supported by grants from National Basic Research Program of China (973 Program, 2010CB529800), the National Natural Science Foundation of China (NSFC-Yunnan jointed funding U1132601) and the grant 2011C1139 to Yun Zhang. |
| 收录类别 | SCI |
| 语种 | 英语 |
| 源URL | [http://159.226.149.26:8080/handle/152453/9583]  |
| 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.Key Laboratory of Animal Models and Human Disease Mechanisms, Kunming Institute of Zoology, Chinese Academy of Sciences, Kunming 650223, Yunnan, China 2.The Key Laboratory of Chemistry for Natural Products, Guizhou Province and Chinese Academy of Sciences, Guiyang 550002, China 3.The Graduate School of the Chinese Academy of Sciences, Beijing 100049, China |
| 推荐引用方式 GB/T 7714 | Zeng L,Sun QY,Zhang Y,et al. Molecular cloning and characterization of a complement-depleting factor from king cobra, Ophiophagus hannah[J]. TOXICON,2012,60(3):290-301. |
| APA | Zeng L,Sun QY,Zhang Y,Jin Y,Zhang Y[*],&Lee WH.(2012).Molecular cloning and characterization of a complement-depleting factor from king cobra, Ophiophagus hannah.TOXICON,60(3),290-301. |
| MLA | Zeng L,et al."Molecular cloning and characterization of a complement-depleting factor from king cobra, Ophiophagus hannah".TOXICON 60.3(2012):290-301. |
入库方式： OAI收割
来源：昆明动物研究所
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