Localization of pNT22 70 kDa Heat Shock Cognate-like Protein in the Plasma Membrane

Cell Structure and Function

Weimin Qi¹  Kokichi Kikuchi¹  Seiji Ohtani¹  Toshihiko Torigoe¹  Noriyuki Sato¹  Itaru Hirai¹ 

[1] Department of Pathology, Sapporo Medical University School of Medicine

关键词: hsc73;    cell surface expression;    Triton X-114 phase separation;    epitope mapping;   

DOI  :  10.1247/csf.23.153

学科分类：分子生物学，细胞生物学和基因

来源: Japan Society for Cell Biology

【 摘 要 】

References(27)Cited-By(5)It has been argued that 70 kDa heat shock cognate (hsc73)-like molecules may be expressed on the surface of certain cells, but direct evidence of this has yet to be found. To clarify whether this molecule belongs to hsc73 itself, the membrane protein fraction of Daudi cells was isolated by Triton X-114 phase separation and the reactivity of this membrane protein fraction was assessed with monoclonal antibodies (mAbs) which react with 70 kDa heat shock protein (hsp) family, i.e., NT22, A15 and 3A3. In western blotting analysis, mAb NT22-deflned protein (pNT22) was clearly detected as a membrane protein of Daudi cells with an approximate molecular size of 70 kDa, whereas pNT22 was not recognized by anti-cytoplasmic hsc73/hsp72 mAbs A15 or 3A3. By using deleted recombinant hsc73 proteins, it was determined that mAb NT22 recognizes the N-terminal 350-372 ami no acid stretches of the hsc73 protein. mAb NT22 also reacted with the cell surface protein of Daudi cells in FACS analysis. Taken together, our present data strongly suggest that pNT22 may be a novel hsc73-like protein that is localized in the plasma membrane.

【 授权许可】

Unknown