Utility of formaldehyde cross-linking and mass spectrometry in the study of protein-protein interactions
Utility of formaldehyde cross-linking and mass spectrometry in the study of protein-protein interactions
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DOI：
10.1002/jms.1415
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发表时间：
2008-06-01
影响因子：
2.3
通讯作者：
Kast, Juergen
中科院分区：
文献类型：
--
作者：
Sutherland, Brent W.;Toews, Judy;Kast, Juergen
几十年来，甲醛一直被常规用于交联细胞、组织中的蛋白质，在某些情况下，甚至是整个生物体。由于其体积小，甲醛可以很容易地渗透细胞壁和膜，导致有效的交联，即蛋白质，DNA和其他反应分子之间形成共价键。事实上，甲醛交联是许多主流分析/细胞生物学技术的工具组分，包括细胞核中发现的蛋白质-DNA复合物的染色质免疫沉淀（ChIP）;细胞、组织和器官内蛋白质表达和定位的免疫组织学分析;以及用于可视化聚丙烯酰胺凝胶中低丰度蛋白质的质谱（MS）兼容银染方法。然而，尽管甲醛非常适合用于分析细胞内的蛋白质环境，但它还没有被普遍用于蛋白质-蛋白质相互作用和细胞网络的直接分析。本文的总体目的是讨论最近的进展，在使用甲醛交联结合MS为基础的方法。使用甲醛的关键优势和局限性，目前用于研究蛋白质-蛋白质相互作用的其他交联剂和技术的突出，和基于甲醛的实验方法，被证明是非常有前途的能力，准确和有效地识别新的蛋白质-蛋白质和多蛋白质相互作用复合物。版权所有（c）2008约翰威利父子有限公司。
For decades, formaldehyde has been routinely used to cross-link proteins in cells, tissue, and in some instances, even entire organisms. Due to its small size, formaldehyde can readily permeate cell walls and membranes, resulting in efficient cross-linking, i.e. the formation of covalent bonds between proteins, DNA, and other reactive molecules. Indeed, formaldehyde cross-linking is an instrumental component of many mainstream analytical/cell biology techniques including chromatin immunoprecipitation (ChIP) of protein-DNA complexes found in nuclei; immunohistological analysis of protein expression and localization within cells, tissues, and organs; and mass spectrometry (MS)-compatible silver-staining methodologies used to visualize low abundance proteins in polyacrylamide gels. However, despite its exquisite suitability for use in the analysis of protein environments within cells, formaldehyde has yet to be commonly employed in the directed analysis of protein-protein interactions and cellular networks. The general purpose of this article is to discuss recent advancements in the use of formaldehyde cross-linking in combination with MS-based methodologies. Key advantages and limitations to the use of formaldehyde over other cross-linkers and technologies currently used to study protein-protein interactions are highlighted, and formaldehyde-based experimental approaches that are proving very promising in their ability to accurately and efficiently identify novel protein-protein and multiprotein interaction complexes are presented. Copyright (c) 2008 John Wiley & Sons, Ltd.