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Mitochondrial oxidative phosphorylation system is recruited to detergent‐resistant lipid rafts during myogenesis
Author(s) -
Kim BongWoo,
Lee JoongWon,
Choo HyoJung,
Lee Chang Seok,
Jung SoonYoung,
Yi JaeSung,
Ham YoungMi,
Lee JooHyung,
Hong Jin,
Kang MinJu,
Chi SungGil,
Hyung SeokWon,
Lee SangWon,
Kim Hwan Myung,
Cho Bong Rae,
Min DoSik,
Yoon Gyesoon,
Ko YoungGyu
Publication year - 2010
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.200900826
Subject(s) - lipid raft , myogenesis , oxidative phosphorylation , sarcolemma , microbiology and biotechnology , biology , biochemistry , skeletal muscle , myocyte , chemistry , signal transduction , endocrinology
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.