Purification of protein complexes by immunoaffinity chromatography: application to transcription machinery.

Purification of protein complexes by immunoaffinity chromatography: application to transcription machinery.
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通过免疫亲和层析纯化蛋白质复合物:在转录机器中的应用。

DOI:
10.1007/0-387-25856-6_6
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发表时间:
2006
期刊:
Genetic engineering.
影响因子:
--
通讯作者:
Burgess,RichardR
Burgess,RichardR
中科院分区:
--
文献类型:
--
作者:
Thompson,NancyE;Jensen,DebraBridges;Lamberski,JenniferA;Burgess,RichardR

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许多蛋白质必须与其他蛋白质相互作用才能实现细胞和亚细胞结构、获得生物活性和/或调节生物活性。传统上,蛋白质纯化方法常常为了“纯度”而分离蛋白质复合物,或者由于分离过程的化学或物理基础而无意中破坏了复合物。有时可以通过组合单独纯化的蛋白质来重建复合物,但在许多情况下,活性无法恢复。此外,一旦结合伙伴物理上分开,通常很难识别它们。本章将解决纯化蛋白质复合物的挑战,并将特别关注实现这种纯化的免疫亲和层析方法。蛋白质的免疫亲和纯化利用了抗体对抗原的高特异性。这种强大的方法可以在一个步骤中将蛋白质纯化 1,000 倍。一般来说,抗原是
Many proteins must interact with other proteins to achieve cellular and subcellular structures, to acquire biological activity, and/or to regulate biological activity. Traditionally, protein purification methods have often resulted in the separation of protein complexes for the sake of “purity” or have inadvertently destroyed complexes as a result of the chemical or physical basis of the separation procedure. Sometimes the complex can be reconstituted by combining individually purified proteins, but, in many cases, activity cannot be recovered. In addition, it is often difficult to identify the binding partners once they are physically separated. This chapter will address the challenge of purifying protein complexes and will specifically concentrate on the method of immunoaffinity chromatography to achieve this purification. Immunoaffinity purification of proteins takes advantage of the high specificity of an antibody for an antigen. This powerful method can result in a 1,000-fold purification of a protein in a single step. Generally, the antigen is the
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