Mitochondrial oxidative phosphorylation system is recruited to detergent-resistant lipid rafts during myogenesis

Bong Woo Kim, Joong Won Lee, Hyo Jung Choo, Chang Seok Lee, Soon Young Jung, Jae Sung Yi, Young Mi Ham, Joo Hyung Lee, Jin Hong, Min Ju Kang, Sung Gil Chi, Seok Won Hyung, Sang Won Lee, Hwan Myung Kim, Bong Rae Cho, Do Sik Min, Gyesoon Yoon, Young Gyu Ko

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

Since detergent-resistant lipid rafts play important roles in the signal transduction for myogenesis, their comprehensive proteomic analysis could provide new insights to understand their function in myotubes. Here, the detergent-resistant lipid rafts were isolated from C2C12 myotubes and analyzed by capillary RPLC/MS/MS. Among the 327 proteins (or protein groups) identified, 28% were categorized to the plasma membrane or raft proteins, 29% to mitochondria, 20% to microsomal proteins, 10% to other proteins, and 13% to unknown proteins. The localization of oxidative phosphorylation (OXPHOS) complexes in the sarcolemma lipid rafts was further confirmed from C2C12 myotubes by cellular fractionation, surface-biotin labeling, immunofluorescence, and lipid raft fractionation. After adding exogenous cytochrome c, the sarcolemma isolated from myotubes had an ability to consume oxygen in the presence of NADH or succinate. The generation of NADH-dependent extracellular superoxide was increased by inhibiting or downregulating OXPHOS I, III, and IV in myotubes, indicating that OXPHOS proteins are major sources for extracellular ROS in skeletal muscle. With all these data, we can conclude that OXPHOS proteins are associated with the sarcolemma lipid rafts during C2C12 myogenesis to generate extracellular ROS.

Original languageEnglish
Pages (from-to)2498-2515
Number of pages18
JournalProteomics
Volume10
Issue number13
DOIs
Publication statusPublished - 2010 Jul

Keywords

  • Capillary RPLC/MS/MS
  • Cell biology
  • Lipid rafts
  • Myogenesis
  • Oxidative phosphorylation proteins
  • Respiration

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

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