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蛇毒蛋白质组学中的一种蛋白质解聚策略

A Protein Decomplexation Strategy in Snake Venom Proteomics.

作者信息

Tan Choo Hock, Tan Kae Yi, Tan Nget Hong

机构信息

Venom Research and Toxicology Laboratory, Department of Pharmacology, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia.

Protein and Interactomic Laboratory, Department of Molecular Medicine, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia.

出版信息

Methods Mol Biol. 2019;1871:83-92. doi: 10.1007/978-1-4939-8814-3_5.

Abstract

Snake venoms are complex mixtures of proteins and peptides that play vital roles in the survival of venomous snakes. As with their diverse pharmacological activities, snake venoms can be highly variable, hence the importance of understanding the compositional details of different snake venoms. However, profiling venom protein mixtures is challenging, in particular when dealing with the diversity of protein subtypes and their abundances. Here we described an optimized strategy combining a protein decomplexation method with in-solution trypsin digestion and mass spectrometry of snake venom proteins. The approach involves the integrated use of C reverse-phase high-performance liquid chromatography (RP-HPLC), sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and nano-electrospray ionization tandem mass spectrometry (nano-ESI-LC-MS/MS).

摘要

蛇毒是蛋白质和肽的复杂混合物,在毒蛇的生存中起着至关重要的作用。与其多样的药理活性一样,蛇毒可能具有高度变异性,因此了解不同蛇毒的组成细节非常重要。然而,对毒液蛋白质混合物进行分析具有挑战性,尤其是在处理蛋白质亚型的多样性及其丰度时。在此,我们描述了一种优化策略,该策略将蛋白质解聚方法与蛇毒蛋白质的溶液内胰蛋白酶消化和质谱分析相结合。该方法涉及综合使用C18反相高效液相色谱(RP-HPLC)、十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)和纳米电喷雾电离串联质谱(nano-ESI-LC-MS/MS)。

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