Fluorescence-Based Assay For Measuring OMA1 Activity

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.

Keywords: Fluorescence-based reporter assay; Fusion; Mitochondria; OMA1; Protease.

MeSH terms

  • Cells, Cultured
  • Enzyme Assays / methods*
  • Fluorescent Dyes / chemistry*
  • GTP Phosphohydrolases / metabolism*
  • Humans
  • Metalloendopeptidases / metabolism*
  • Mitochondria / enzymology*
  • Mitochondrial Dynamics
  • Peptides / chemistry*

Substances

  • Fluorescent Dyes
  • Peptides
  • Metalloendopeptidases
  • molecule metalloprotease-related protein-1, human
  • GTP Phosphohydrolases
  • OPA1 protein, human