Lipid raft proteome reveals ATP synthase complex in the cell surface

Tae Jung Bae, Min Sik Kim, Jun Woo Kim, Bong Woo Kim, Hyo Jung Choo, Joong Won Lee, Ki Bum Kim, Seok Lee Chang, Ji Hyun Kim, Young Chang Sun, Chang Yuil Kang, Sang-Won Lee, Young-Gyu Ko

Research output: Contribution to journalArticle

137 Citations (Scopus)

Abstract

Since detergent-resistant lipid rafts are involved in pathogen invasion, cholesterol homeostasis, angiogenesis, neurodegenerative diseases and signal transduction, protein identification in the rafts could provide important information to study their function. Here, we analyzed detergent-resistant raft proteins isolated from rat liver by capillary liquid chromatography-tandem mass spectrometry. Out of 196 proteins identified, 32% belonged to the raft or plasma membrane, 24% to mitochondrial, 20% to microsomal, 7% to miscellaneous, and 17% are unknown proteins. For example, membrane-bound receptors, trimeric GTP-binding proteins, ATP-binding cassette transporters, and glycosylphosphatidylinositol-anchored proteins were identified in this analysis. Unexpectedly, there were many mitochondrial proteins, raising a new issue for the presence of mitochondrial rafts or the localization of mitochondrial proteins into plasma membrane rafts. We confirmed that ATP synthase α and β were expressed on the surface of the plasma membrane in HepG2 hepatocytes by immunofluorescence, cell surface biotinylation, and cellular fractionation. They had two distinct biochemical properties, detergent insolubility and low density, suggesting that the ATP synthase complex might be located in plasma membrane rafts as well as in the mitochondria.

Original languageEnglish
Pages (from-to)3536-3548
Number of pages13
JournalProteomics
Volume4
Issue number11
DOIs
Publication statusPublished - 2004 Nov 1

Keywords

  • ATP synthase
  • Capillary liquid chromatography-tandem mass spectrometry
  • Lipid rafts
  • Mitochondria

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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  • Cite this

    Bae, T. J., Kim, M. S., Kim, J. W., Kim, B. W., Choo, H. J., Lee, J. W., Kim, K. B., Chang, S. L., Kim, J. H., Sun, Y. C., Kang, C. Y., Lee, S-W., & Ko, Y-G. (2004). Lipid raft proteome reveals ATP synthase complex in the cell surface. Proteomics, 4(11), 3536-3548. https://doi.org/10.1002/pmic.200400952