Macromolecular affinity labeling.
Macromolecular affinity labeling.
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大分子亲和标记。
DOI：
10.1007/bf02623719
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发表时间：
1989
期刊：
影响因子：
--
通讯作者：
Ji,I
中科院分区：
文献类型：
--
作者：
Ji,TH;Ji,I
自从首次描述大分子亲和标记方法(Ji，1976)以来，许多结合蛋白已被成功地用光亲和标记(Ji，1976)和亲和交联法(Pilch和捷克，1979)鉴定出来。这些大分子亲和标记方法现已成为受体研究的标准方法。它们的应用已经超越了结合蛋白的鉴定，扩展到阐明各种途径、内吞过程和蛋白质之间的相互作用。预测分子相互作用部位的其他方法是使用抗体和重组突变分子。然而，与这些方法不同的是，亲和标记法不仅能够识别相互作用的位点，而且还能够将配体中的位点与受体中的相应位点进行匹配。试剂的改进和方法的灵敏度使检测低浓度表达的受体成为可能，例如每个细胞只有几百个分子。最近描述了循序渐进的方案(纪万昌等人，1989)。在这篇文章中，我将讨论大分子光亲和标记和亲和交联应用的最新进展。
Since the first description of the macromolecular affinity labeling methods (Ji, 1976) numerous binding proteins have successfully been identified by photoaffini ty labeling (Ji, 1976) and affinity crosslinking (Pilch and Czech, 1979). These macromolecular affinity labeling methods are now established as a standard approach for receptor studies. Their applications have expanded beyond the identification of binding proteins to the elucidation of various pathways, the process of endocyto sis, and interactions of the proteins. Other ways to predict the interacting sites of a molecule are the use of antibodies and recombinant mutant molecules. However, unlike these methods, affinity labeling is not only capable of identifying the interacting sites but also of matching the site in the ligand to the corresponding site in the receptor. Improvements in the reagents and the sensitivity of the methods have made possible the examination of receptors expressed at low concentra tions, such as a few hundred molecules per a cell. Step-by-step protocols have recently been described (Ji et al., 1989). In this article I will discuss the recent developments in the application of macromolecular photoaffinity labeling and affinity crosslinking.