Proteome profile of functional mitochondria from human skeletal muscle using one-dimensional gel electrophoresis and HPLC-ESI-MS/MS

Natalie Lefort, Zhengping Yi, Benjamin Bowen, Brian Glancy, Eleanna A. De Filippis, Rebekka Mapes, Hyonson Hwang, Charles R. Flynn, Wayne T. Willis, Anthony Civitarese, Kurt Højlund, Lawrence J. Mandarino

Research output: Contribution to journalArticle

52 Scopus citations

Abstract

Mitochondria can be isolated from skeletal muscle in a manner that preserves tightly coupled bioenergetic function in vitro. The purpose of this study was to characterize the composition of such preparations using a proteomics approach. Mitochondria isolated from human vastus lateralis biopsies were functional as evidenced by their response to carbohydrate and fat-derived fuels. Using one-dimensional gel electrophoresis and HPLC-ESI-MS/MS, 823 unique proteins were detected, and 487 of these were assigned to the mitochondrion, including the newly characterized SIRT5, MitoNEET and RDH13. Proteins detected included 9 of the 13 mitochondrial DNA-encoded proteins and 86 of 104 electron transport chain (ETC) and ETC-related proteins. In addition, 59 of 78 proteins of the 55S mitoribosome, several TIM and TOM proteins and cell death proteins were present. This study presents an efficient method for future qualitative assessments of proteins from functional isolated mitochondria from small samples of healthy and diseased skeletal muscle.

Original languageEnglish (US)
Pages (from-to)1046-1060
Number of pages15
JournalJournal of Proteomics
Volume72
Issue number6
DOIs
Publication statusPublished - Aug 20 2009

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Keywords

  • HPLC-ESI-MS/MS
  • Human skeletal muscle
  • Mitochondria
  • One-dimensional-gel electrophoresis
  • Proteome

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics

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