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印度东部圆斑蝰蛇(Daboia russelii)毒液的蛋白质组分析和免疫分析:RVV 组成与 RV 咬伤后临床症状的相关性。

Proteomic Analysis and Immuno-Profiling of Eastern India Russell's Viper ( Daboia russelii) Venom: Correlation between RVV Composition and Clinical Manifestations Post RV Bite.

机构信息

Microbial Biotechnology and Protein Research Laboratory, Department of Molecular Biology and Biotechnology , Tezpur University , Tezpur 784028 , Assam , India.

出版信息

J Proteome Res. 2018 Aug 3;17(8):2819-2833. doi: 10.1021/acs.jproteome.8b00291. Epub 2018 Jul 10.

Abstract

The proteomes of Russell's viper venom (RVV) from Burdwan (RVV B) and Nadia (RVV N), the two districts of West Bengal, eastern India (EI), were investigated by gel-filtration chromatography (GFC) followed by tandem mass spectrometry of tryptic fragments of the fractions. A total of 73 and 69 proteins belonging to 15 snake venom protein families were identified in RVV B and RVV N, respectively, by MS/MS search against Viperidae (taxid 8689) protein entries of the nonredundant NCBI database. The minor differences in venom composition of both the EI RV were established unequivocally by their biochemical and pharmacological properties and by SDS-PAGE, gel filtration chromatography, and LC-MS/MS analyses. The composition of EI RVVs was well correlated with published reports on the pathophysiology of RV-envenomed patients from this part of the country. Venom-antivenom cross-reactivity determined by ELISA, Western blotting, and antivenomics approaches demonstrated poor recognition of low molecular mass (<20 kDa) RVV proteins by commercial polyvalent antivenoms, which was substantiated by neutralization of RVV enzymes by antivenom.

摘要

本研究采用凝胶过滤色谱(GFC)分离结合肽段的串联质谱分析,对产自印度东部西孟加拉邦伯德万(RVV B)和那迪亚(RVV N)两个地区的圆斑蝰蛇毒(RVV)的蛋白质组进行了分析。通过与非冗余 NCBI 数据库中蝰科(taxid 8689)蛋白条目进行 MS/MS 搜索,在 RVV B 和 RVV N 中分别鉴定出了 73 种和 69 种属于 15 种蛇毒蛋白家族的蛋白质。这两种来自印度东部的圆斑蝰蛇毒的生物化学和药理学特性以及 SDS-PAGE、凝胶过滤色谱和 LC-MS/MS 分析,明确证实了它们在毒液组成上的细微差异。来自该地区的 RV 蛇毒中毒患者的病理生理学研究报告与 EI-RVV 的组成情况非常吻合。ELISA、Western blot 和抗蛇毒组学方法确定的毒液-抗毒液交叉反应性表明,商业多价抗蛇毒血清对低分子量(<20 kDa)RVV 蛋白的识别能力较差,抗蛇毒血清能中和 RVV 酶,这一结果得到了证实。

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