Assessing Protein Interactions for Clustering of Mitochondrial Urea Cycle Enzymes.
Assessing Protein Interactions for Clustering of Mitochondrial Urea Cycle Enzymes.
复制标题
评估线粒体尿素循环酶聚类的蛋白质相互作用。
DOI：
10.1007/978-1-0716-2269-8_5
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发表时间：
2022
期刊：
影响因子：
--
通讯作者：
Jaiswal,Jyoti
中科院分区：
文献类型：
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作者：
Caldovic,Ljubica;Bhuvanendran,Shivaprasad;Jaiswal,Jyoti
酶簇是一种涉及在共同的亚细胞或亚细胞器区室中一起发挥作用的蛋白质的划分的现象。传统的遗传、生物化学和生物物理方法用于研究具有确定化学计量的复合物中蛋白质-蛋白质相互作用，当应用于簇状蛋白质时，会产生不确定的结果。本章介绍了研究聚集蛋白的方法组合，包括免疫共沉淀、纯化线粒体中的生化共定位以及内源蛋白原位超分辨率成像。这些方法可用于研究形成簇的蛋白质之间的相互作用。我们将通过使用位于线粒体基质中并在线粒体内膜上形成簇的尿素循环酶来说明这种方法。
Enzyme clustering is a phenomenon that involves partitioning of proteins that function together in a common subcellular or sub-organellar compartment. Traditional genetic, biochemical, and biophysical approaches for studying protein–protein interactions in complexes with defined stoichiometry yield inconclusive results when applied to clustered proteins. This chapter describes a combination of approaches to study clustered proteins including co-immunoprecipitation, biochemical co-localization in purified mitochondria, and super resolution imaging of endogenous proteins in situ. These approaches can be used to study interactions among proteins that form clusters. We will illustrate this approach by using the urea cycle enzymes that localize in the mitochondrial matrix, and form clusters at the inner mitochondrial membrane.