A 60 kDa Plasma Membrane Protein Changes its Localization to Autophagosome and Autolysosome Membranes during Induction of Autophagy in Rat Hepatoma Cell Line, H-4-II-E Cells

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We previously reported the preparation and characterization of an antibody against membrane fraction of autolysosomes from rat liver (J. Histochem. Cytochem. 38, 1571-1581, 1990). Immunoblot analyses of total membrane fraction of a rat hepatoma cell line, H-4-II-E cells by this antibody suggested that H-4-II-E cells expressed several autolysosomal proteins, including a protein with apparent molecular weight of 60 kDa. It was suggested that this 60 kDa protein was a peripheral membrane protein, because it was eluted from the membrane by sodium carbonate treatment. We prepared an antibody against this 60 kDa protein by affinity purification method, and examined its behavior during induction of autophagy. Autophagy was induced by transferring the cells from Dulbecco's modified Eagle medium(DMEM) containing 12% fetal calf serum into Hanks' balance salt solution. In DMEM, the 60 kDa protein showed diffused immunofluorescence pattern, and immunoelectron microscopy suggested that this protein was located on the extracellular side of the plasma membrane. After inducing autophagy, the immunofluorescence configuration of the 60 kDa protein changed from the diffused pattern to a granulous one. Immunoelectron microscopy showed that the 60 kDa protein was localized on the luminal side of the limiting membrane of autolysosomes and endosomes. in the presence of bafilomycin A<sub>1</sub> which prevents fusion between autophagosomes and lysosomes, the 60 kDa protein was localized on the limiting membrane of the autophagosomes and endosomes. These results suggest that the 60 kDa protein is transported from the plasma membrane to the autophagosome membrane through the endosomes.

収録刊行物

  • Cell structure and function

    Cell structure and function 24(2), 59-70, 1999-04

    Japan Society for Cell Biology

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各種コード

  • NII論文ID(NAID)
    10002420728
  • NII書誌ID(NCID)
    AA0060007X
  • 本文言語コード
    ENG
  • 資料種別
    ART
  • ISSN
    03867196
  • NDL 記事登録ID
    4728475
  • NDL 雑誌分類
    ZR1(科学技術--生物学)
  • NDL 請求記号
    Z53-V38
  • データ提供元
    CJP書誌  CJP引用  NDL  J-STAGE 
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