检测热休克蛋白 90 同工型的翻译后修饰。

Detecting Posttranslational Modifications of Hsp90 Isoforms.

机构信息

Department of Urology, SUNY Upstate Medical University, Syracuse, NY, USA.

Urologic Oncology Branch, Center for Cancer Research, National Cancer Institute, Bethesda, MD, USA.

出版信息

Methods Mol Biol. 2023;2693:125-139. doi: 10.1007/978-1-0716-3342-7_11.

DOI:10.1007/978-1-0716-3342-7_11

PMID:37540432

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10518168/

Abstract

The molecular chaperone heat shock protein 90 (Hsp90) is essential in eukaryotes. Hsp90 chaperones proteins that are important determinants of multistep carcinogenesis. There are multiple Hsp90 isoforms including the cytosolic Hsp90α and Hsp90β as well as GRP94 located in the endoplasmic reticulum and TRAP1 in the mitochondria. The chaperone function of Hsp90 is linked to its ability to bind and hydrolyze ATP. Co-chaperones and posttranslational modifications (such as phosphorylation, SUMOylation, and ubiquitination) are important for Hsp90 stability and regulation of its ATPase activity. Both mammalian and yeast cells can be used to express and purify Hsp90 and TRAP1 and also detect post-translational modifications by immunoblotting.

摘要

热休克蛋白 90（Hsp90）是真核生物中必需的分子伴侣。Hsp90 伴侣蛋白是多步骤致癌作用的重要决定因素。有多种 Hsp90 同工型，包括胞质 Hsp90α 和 Hsp90β 以及内质网中的 GRP94 和线粒体中的 TRAP1。Hsp90 的伴侣功能与其结合和水解 ATP 的能力有关。共伴侣和翻译后修饰（如磷酸化、SUMO 化和泛素化）对于 Hsp90 的稳定性和其 ATP 酶活性的调节很重要。哺乳动物和酵母细胞都可用于表达和纯化 Hsp90 和 TRAP1，并通过免疫印迹检测翻译后修饰。