基于荧光的 OMA1 活性检测方法。

Fluorescence-Based Assay For Measuring OMA1 Activity.

机构信息

Department of Pharmacology and Toxicology, University of Arkansas for Medical Sciences, Little Rock, AR, USA.

出版信息

Methods Mol Biol. 2021;2276:325-332. doi: 10.1007/978-1-0716-1266-8_24.

Abstract

Mitochondrial fusion depends on proteolytic processing of the dynamin-related GTPase protein, OPA1, which is regulated by the mitochondrial zinc metalloproteinase, OMA1. Last year we published a report describing a novel approach to directly measure the enzymatic activity of OMA1 in whole cell lysates. This fluorescence-based reporter assay utilizes an eight amino acid peptide sequence referred to as the S1 cleavage site where OMA1 cleaves within OPA1 and is flanked by a fluorophore and quencher. In this chapter, we provide additional insight into the OMA1 activity assay.

摘要

线粒体融合依赖于解旋酶相关 GTP 酶蛋白 OPA1 的蛋白水解加工，其受线粒体锌金属蛋白酶 OMA1 调控。去年，我们发表了一篇报道，描述了一种直接测量全细胞裂解物中 OMA1 酶活性的新方法。该荧光报告测定法利用了一个八氨基酸肽序列，称为 S1 切割位点，在该位点 OMA1 在 OPA1 内切割，其两侧是荧光团和猝灭剂。在本章中，我们将提供对 OMA1 活性测定的更多见解。