利用蛋白质亲和纯化技术从秀丽隐杆线虫中分离丝氨酸蛋白酶抑制剂相互作用蛋白。
Isolation of serpin-interacting proteins in C. elegans using protein affinity purification.
机构信息
Department of Pediatrics, Cell Biology and Physiology, University of Pittsburgh School of Medicine, Children's Hospital of Pittsburgh of UPMC, and Magee-Womens Hospital of UPMC, Pittsburgh, PA 15224, USA.
Biomedical Mass Spectrometry Center, University of Pittsburgh Schools of the Health Sciences, Pittsburgh, PA 15213, USA.
出版信息
Methods. 2014 Aug 1;68(3):536-41. doi: 10.1016/j.ymeth.2014.04.019. Epub 2014 May 2.
Caenorhabditis elegans is a useful model organism for combining multiple imaging, genetic, and biochemical methodologies to gain more insight into the biological function of specific proteins. Combining both biochemical and genetic analyses can lead to a better understanding of how a given protein may function within the context of a network of other proteins or specific pathway. Here, we describe a protocol for the biochemical isolation of serpin-interacting proteins using affinity purification and proteomic analysis. As the knowledge of in vivo serpin interacting partners in C. elegans has largely been obtained using genetic and in vitro recombinant protein studies, this protocol serves as a complementary approach to provide insight into the biological function and regulation of serpins.
秀丽隐杆线虫是一种有用的模式生物，可将多种成像、遗传和生化方法结合起来，深入了解特定蛋白质的生物学功能。将生化和遗传分析结合起来，可以更好地理解给定蛋白质在其他蛋白质网络或特定途径中的功能。本文描述了使用亲和纯化和蛋白质组学分析从秀丽隐杆线虫中生化分离丝氨酸蛋白酶抑制剂相互作用蛋白的方案。由于体内秀丽隐杆线虫丝氨酸蛋白酶抑制剂相互作用伙伴的知识主要是通过遗传和体外重组蛋白研究获得的，因此该方案是一种补充方法，可以深入了解丝氨酸蛋白酶抑制剂的生物学功能和调节。
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