A cell death assay for assessing the mitochondrial targeting of proteins

Daniel Camara Teixeira, Elizabeth L. Cordonier, Subhashinee S.K. Wijeratne, Patricia Huebbe, Augusta Jamin, Sarah Jarecke, Matthew Wiebe, Janos Zempleni

Research output: Contribution to journalArticlepeer-review

Abstract

The mitochondrial proteome comprises 1000 to 1500 proteins, in addition to proteins for which the mitochondrial localization is uncertain. About 800 diseases have been linked with mutations in mitochondrial proteins. We devised a cell survival assay for assessing the mitochondrial localization in a high-throughput format. This protocol allows us to assess the mitochondrial localization of proteins and their mutants, and to identify drugs and nutrients that modulate the mitochondrial targeting of proteins. The assay works equally well for proteins directed to the outer mitochondrial membrane, inner mitochondrial membrane mitochondrial and mitochondrial matrix, as demonstrated by assessing the mitochondrial targeting of the following proteins: carnitine palmitoyl transferase 1 (consensus sequence and R123C mutant), acetyl-CoA carboxylase 2, uncoupling protein 1 and holocarboxylase synthetase. Our screen may be useful for linking the mitochondrial proteome with rare diseases and for devising drug- and nutrition-based strategies for altering the mitochondrial targeting of proteins.

Original languageEnglish (US)
Pages (from-to)48-54
Number of pages7
JournalJournal of Nutritional Biochemistry
Volume56
DOIs
StatePublished - Jun 2018

Keywords

  • 3-chlorogenic acid
  • Acetyl-CoA carboxylase
  • High-throughput screen
  • Mitochondria
  • Proteome

ASJC Scopus subject areas

  • Endocrinology, Diabetes and Metabolism
  • Biochemistry
  • Molecular Biology
  • Nutrition and Dietetics
  • Clinical Biochemistry

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