蛋白质结构与相互作用研究新方法——交联质谱技术

蛋白质的空间结构信息以及蛋白质间的相互作用信息对于研究蛋白质的功能有重要意义.研究蛋白质结构与相互作用的传统技术,如核磁共振技术、X射线晶体衍射技术等,对于蛋白质的纯度、结晶性和绝对量均有比较高的要求,限制了其广泛应用.交联质谱技术是近十多年来发展起来的新技术,它将质谱技术与交联技术相结合,在研究蛋白质结构与相互作用方面具有速度快、成本小、蛋白质各方面性状要求低等优势.本文就交联质谱技术各个环节的技术方法加以综述,包括交联质谱实验分离富集技术、常见交联剂特性、交联质谱数据库搜索算法、结果验证研究和交联质谱技术的应用等方面,并展望了该研究方向未来的发展.

A New Approach to Protein Structure and Interaction Research: Chemical Cross-linking in Combination With Mass Spectrometry

For any protein, its 3-D structure and interaction with other proteins are vital information for the understanding of its function. Traditionally, NMR and X-ray crystallography are employed to acquire such information at high resolution. However, application of these methods is limited because they are extremely demanding for protein purity and quantity, or protein size and crystalizability. Chemical cross-linking in combination with mass spectrometry (CXMS) is a novel technology developed over the past decade that complements the traditional methods with its many advantages such as high throughput, low cost and relaxed sample requirements. Here we review the various aspects of CXMS, including separation and enrichment techniques, characteristics of common cross-linkers, development of database-search tools for CXMS data, algorithms for validation of results, and application. The future development of CXMS is also discussed.