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GRP78/BiP is Involved in Ouabain-induced Endocytosis of the Na/K-ATPase in LLC-PK1 Cells

Kesiry, Riad

Abstract Details

2004, Master of Science in Biomedical Sciences (MSBS), University of Toledo, Graduate School.
We have demonstrated that ouabain causes dose- and time-dependent decreases both in 86Rb uptake and plasmalemmal Na/K-ATPase content of LLC-PK1 cells (Liu et al., 2002). This phenomenon is related to ouabain-induced endocytosis of plasmalemmal Na/K-ATPase in LLC-PK1 Cells by a clathrin-dependent mechanism (Liu et al., 2004). GRP78/BiP is a resident protein of the endoplasmic reticulum (ER) and acts as a molecular chaperone. Recently, several studies have shown GRP78/BiP is also expressed on the cell surface and forms heterogeneous, high molecular weight complexes with other proteins. To identify the proteins that are possibly involved in ouabain-induced endocytosis of the Na/K-ATPase in LLC-PK1 cells, we separated and identified endosomal proteins by 2D gel electrophoresis and MS/MS from both control and ouabain-treated LLC-PK1 cells. GRP78/BiP was identified by MS/MS as one of several up-regulated proteins and confirmed by Western Blot. Interestingly, GRP78/BiP was significantly up-regulated in both early and late endosomes in response to ouabain treatment. By using a cell surface protein biotinylation technique to isolate the cell surface membrane proteins, we found that GRP/BiP is expressed on the cell surface of LLC-PK1 cells and down-regulated in a time-dependent manner in response to ouabain. By comparing the cellular redistribution, our data suggest that both the Na/K-ATPase a-1 subunit and GRP78/BiP follow the same pattern in response to ouabain.
Joseph Shapiro, M.D. (Advisor)
30 p.

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Citations

  • Kesiry, R. (2004). GRP78/BiP is Involved in Ouabain-induced Endocytosis of the Na/K-ATPase in LLC-PK1 Cells [Master's thesis, University of Toledo]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=mco1096302498

    APA Style (7th edition)

  • Kesiry, Riad. GRP78/BiP is Involved in Ouabain-induced Endocytosis of the Na/K-ATPase in LLC-PK1 Cells. 2004. University of Toledo, Master's thesis. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=mco1096302498.

    MLA Style (8th edition)

  • Kesiry, Riad. "GRP78/BiP is Involved in Ouabain-induced Endocytosis of the Na/K-ATPase in LLC-PK1 Cells." Master's thesis, University of Toledo, 2004. http://rave.ohiolink.edu/etdc/view?acc_num=mco1096302498

    Chicago Manual of Style (17th edition)