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通过交联结合质谱法分析蛋白质结构

Analysis of protein structure by cross-linking combined with mass spectrometry.

作者信息

Petrotchenko Evgeniy V, Makepeace Karl A T, Serpa Jason J, Borchers Christoph H

机构信息

University of Victoria - Genome BC Proteomics Centre, University of Victoria, 3101-4464 Markham St, Victoria, BC, 78Z 7X8, Canada.

出版信息

Methods Mol Biol. 2014;1156:447-63. doi: 10.1007/978-1-4939-0685-7_30.

DOI:10.1007/978-1-4939-0685-7_30
PMID:24792007
Abstract

Cross-linking combined with mass spectrometry is a powerful technique to study protein structure. Here, we present an optimized protocol for the preparation, processing, and analysis of a protein sample cross-linked with isotopically coded, affinity-enrichable, and CID-cleavable cross-linker CyanurBiotinDimercaptoPropionylSuccinimide using LC/ESI-MS/MS on a Thermo Scientific Orbitrap mass spectrometer.

摘要

交联结合质谱分析是研究蛋白质结构的一项强大技术。在此，我们展示了一种优化方案，用于制备、处理和分析使用同位素编码、亲和富集且可经碰撞诱导解离（CID）裂解的交联剂氰尿酸生物素二巯基丙酰琥珀酰亚胺交联的蛋白质样品，并在赛默飞世尔科技的Orbitrap质谱仪上使用液相色谱/电喷雾电离串联质谱（LC/ESI-MS/MS）进行分析。

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