In vivo measurement of synthesis rate of individual skeletal muscle mitochondrial proteins

Abdul Jaleel, Kevin R. Short, Yan Asmann, Katherine A. Klaus, Dawn M. Morse, G. Charles Ford, K Sreekumaran Nair

Research output: Contribution to journalArticle

57 Citations (Scopus)

Abstract

Skeletal muscle mitochondrial dysfunction occurs in many conditions including aging and insulin resistance, but the molecular pathways of the mitochondrial dysfunction remain unclear. Presently, no methodologies are available to measure synthesis rates of individual mitochondrial proteins, which limits our ability to fully understand the translational regulation of gene transcripts. Here, we report a methodology to measure synthesis rates of multiple muscle mitochondrial proteins, which, along with large-scale measurements of mitochondrial gene transcripts and protein concentrations, will enable us to determine whether mitochondrial alteration is due to transcriptional or translational changes. The methodology involves in vivo labeling of muscle proteins with L-[ring- 13C 6] phenylalanine, protein purification by two-dimensional gel electrophoresis of muscle mitochondrial fraction, and protein identification and stable isotope abundance measurements by tandem mass spectrometry. Synthesis rates of 68 mitochondrial and 23 non-mitochondrial proteins from skeletal muscle mitochondrial fraction showed a 10-fold range, with the lowest rate for a structural protein such as myosin heavy chain (0.16 ± 0.04%/h) and the highest for a mitochondrial protein such as dihydrolipoamide branched chain transacylase E2 (1.5 ± 0.42%/h). This method offers an opportunity to better define the translational regulation of proteins in skeletal muscle or other tissues.

Original languageEnglish (US)
JournalAmerican Journal of Physiology - Endocrinology and Metabolism
Volume295
Issue number5
DOIs
StatePublished - Nov 2008

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Muscle Proteins
Mitochondrial Proteins
Skeletal Muscle
Proteins
Mitochondrial Genes
Myosin Heavy Chains
Electrophoresis, Gel, Two-Dimensional
Tandem Mass Spectrometry
Phenylalanine
Isotopes
Insulin Resistance
Muscles
Genes

Keywords

  • Protein synthesis
  • Two-dimensional gel electrophoresis

ASJC Scopus subject areas

  • Physiology
  • Physiology (medical)
  • Endocrinology, Diabetes and Metabolism

Cite this

In vivo measurement of synthesis rate of individual skeletal muscle mitochondrial proteins. / Jaleel, Abdul; Short, Kevin R.; Asmann, Yan; Klaus, Katherine A.; Morse, Dawn M.; Ford, G. Charles; Nair, K Sreekumaran.

In: American Journal of Physiology - Endocrinology and Metabolism, Vol. 295, No. 5, 11.2008.

Research output: Contribution to journalArticle

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