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  • 标题:Identification of transmembrane bridging proteins in the plasma membrane of cultured mouse L cells
  • 本地全文:下载
  • 作者:R M Evans ; L M Fink
  • 期刊名称:Proceedings of the National Academy of Sciences
  • 印刷版ISSN:0027-8424
  • 电子版ISSN:1091-6490
  • 出版年度:1977
  • 卷号:74
  • 期号:12
  • 页码:5341-5344
  • DOI:10.1073/pnas.74.12.5341
  • 语种:English
  • 出版社:The National Academy of Sciences of the United States of America
  • 摘要:Studies were carried out to identify transmembrane bridging proteins in the plasma membrane of mouse L-929 cells. Cells grown in suspension culture were 125I-labeled by lactoperoxidase and allowed to ingest latex particles to produce inside-out membrane phagosome preparations. Phagosomes were isolated and the inner membrane surface was labeled with N-(5'-aminopentyl)-5-dimethylamino-1-naphthalenesulfonamide (dansylcadavarine) by a transglutaminase-catalyzed reaction. The phagosome membrane proteins were solubilized and dansylcadavarine-labeled proteins were isolated by anti-dansyl immunoadsorbent affinity chromatography. Dansylcadavarine-labeled proteins were analyzed by sodium dodecyl sulfate/polyacrylamide gel electrophoresis and autoradiography for the presence of 125I-labeled material. By this technique, two iodinated proteins with molecular weights of approximately 50,000 and 80,000 appear to be selectively retained by the anti-dansyl immunoadsorbent, suggesting that these proteins span the plasma membrane.
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