Screening for post-translational modifications in conotoxins using liquid chromatography/mass spectrometry: an important component of conotoxin discovery
Screening for post-translational modifications in conotoxins using liquid chromatography/mass spectrometry: an important component of conotoxin discovery
复制标题
DOI：
10.1016/j.toxicon.2006.01.021
复制
发表时间：
2006-05-01
期刊：
影响因子：
2.8
通讯作者：
Sweedler, Jonathan V.
中科院分区：
文献类型：
--
作者：
Jakubowski, Jennifer A.;Kelley, Wayne P.;Sweedler, Jonathan V.
质谱法因其选择性、灵敏、信息丰富的分析能力而成为芋螺毒素分析的重要技术。使用液相色谱/质谱法，可以对芋螺毒液进行分级，并对肽进行特定翻译后修饰的调查，表明这些毒素成分可能具有重要的生物学功能。以纹状芋螺和胜利芋螺毒液为模型，通过质谱分析过程中观察到的同位素分布和不稳定损失等特征来鉴定溴化、羧化和糖基化修饰。这种修饰筛选方法能够鉴定胜利 C 溴羧基芋螺毒素（命名为 vc5c），作为详细质谱分析的候选毒素。使用cDNA序列结合液相色谱/质谱和纳电喷雾电离-离子阱-质谱，确定vc5c的序列为ICCYPNXWCCD，其中W是6-溴色氨酸，X是γ-羧基谷氨酸，C是二硫键连接的半胱氨酸。这代表了第九种 T 超家族（-CC-CC- 支架）毒素，已从毒液中分离出来并进行了表征。 (c) 2006 Elsevier Ltd. 保留所有权利。
Mass spectrometry has emerged as an important technique for conotoxin analysis due to its capacity for selective, sensitive, information-rich analyses. Using liquid chromatography/mass spectrometry, Conus venom can be fractionated and the peptides surveyed for specific post-translational modifications, indicating those toxin components likely to have an important biological function. With Conus striatus and Conus victoriae venom as models, bromination, carboxylation and glycosylation modifications are identified through characteristics such as isotopic distribution and labile losses observed during mass spectrometric analysis. This modification screening approach enables the identification of a C victoriae bromo-carboxy-conotoxin, designated vc5c, as a candidate for detailed mass spectrometric analysis. Using a cDNA sequence coupled with liquid chromatography/mass spectrometry and nanoelectrospray ionization-ion trap-mass spectrometry, the sequence of vc5c is determined to be ICCYPNXWCCD, where W is 6-bromotryptophan, X is gamma-carboxy glutamate and C is disulfide-linked cysteine. This represents the ninth T-superfamily (-CC-CC- scaffold) toxin that has been isolated from venom and characterized. (c) 2006 Elsevier Ltd. All rights reserved.