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 S Wiebe, Janos Zempleni

Research output: Contribution to journalArticle

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

Fingerprint

Mitochondrial Proteins
Cell death
Assays
Cell Death
Mitochondrial Membranes
Proteome
Proteins
Acetyl-CoA Carboxylase
Carnitine
Consensus Sequence
Protein Transport
Transferases
Rare Diseases
Membranes
Pharmaceutical Preparations
Cell Survival
Mutant Proteins
Nutrition
Nutrients
Food

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

Cite this

Camara Teixeira, D., Cordonier, E. L., Wijeratne, S. S. K., Huebbe, P., Jamin, A., Jarecke, S., ... Zempleni, J. (2018). A cell death assay for assessing the mitochondrial targeting of proteins. Journal of Nutritional Biochemistry, 56, 48-54. https://doi.org/10.1016/j.jnutbio.2018.01.006

A cell death assay for assessing the mitochondrial targeting of proteins. / Camara Teixeira, Daniel; Cordonier, Elizabeth L.; Wijeratne, Subhashinee S.K.; Huebbe, Patricia; Jamin, Augusta; Jarecke, Sarah; Wiebe, Matthew S; Zempleni, Janos.

In: Journal of Nutritional Biochemistry, Vol. 56, 06.2018, p. 48-54.

Research output: Contribution to journalArticle

Camara Teixeira D, Cordonier EL, Wijeratne SSK, Huebbe P, Jamin A, Jarecke S et al. A cell death assay for assessing the mitochondrial targeting of proteins. Journal of Nutritional Biochemistry. 2018 Jun;56:48-54. https://doi.org/10.1016/j.jnutbio.2018.01.006
Camara Teixeira, Daniel ; Cordonier, Elizabeth L. ; Wijeratne, Subhashinee S.K. ; Huebbe, Patricia ; Jamin, Augusta ; Jarecke, Sarah ; Wiebe, Matthew S ; Zempleni, Janos. / A cell death assay for assessing the mitochondrial targeting of proteins. In: Journal of Nutritional Biochemistry. 2018 ; Vol. 56. pp. 48-54.
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