title: Cell surface relocalization of the endoplasmic reticulum chaperone and unfolded protein response regulator GRP78/BiP.
abstract: The recent discovery that GRP78/BiP, a typical endoplasmic reticulum (ER) lumenal chaperone, can be expressed on the cell surface, interacting with an increasing repertoire of surface proteins and acting as receptor in signaling pathways, represents a paradigm shift in its biological function. However, the mechanism of GRP78 trafficking from the ER to the cell surface is not well understood. Using a combination of cellular, biochemical, and mutational approaches, we tested multiple hypotheses. Here we report that ER stress actively promotes GRP78 localization on the cell surface, whereas ectopic expression of GRP78 is also able to cause cell surface relocation in the absence of ER stress. Moreover, deletion of the C-terminal ER retention motif in GRP78 alters its cell surface presentation in a dose-dependent manner; however, mutation of the putative O-linked glycosylation site Thr(648) of human GRP78 is without effect. We also identified the exposure of multiple domains of GRP78 on the cell surface and determined that binding of extracellular GRP78 to the cell surface is unlikely. A new topology model for cell surface GRP78 is presented.
authors.affiliation: Departments of Biochemistry and Molecular Biology, Keck School of Medicine of the University of Southern California, Los Angeles, California 90089-9176.
authors.forename: Yi
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Departments of Pathology, USC Norris Comprehensive Cancer Center, Keck School of Medicine of the University of Southern California, Los Angeles, California 90089-9176.
authors.forename: Ren
authors.initials: R
authors.lastname: Liu
authors.affiliation: Departments of Biochemistry and Molecular Biology, Keck School of Medicine of the University of Southern California, Los Angeles, California 90089-9176.
authors.forename: Min
authors.initials: M
authors.lastname: Ni
authors.affiliation: Departments of Pathology, USC Norris Comprehensive Cancer Center, Keck School of Medicine of the University of Southern California, Los Angeles, California 90089-9176.
authors.forename: Parkash
authors.initials: P
authors.lastname: Gill
authors.affiliation: Departments of Biochemistry and Molecular Biology, Keck School of Medicine of the University of Southern California, Los Angeles, California 90089-9176. Electronic address: amylee@ccnt.usc.edu.
authors.forename: Amy S
authors.initials: AS
authors.lastname: Lee
mesh_terms: D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D004721:Endoplasmic Reticulum / Q000378:metabolism*; D000091342:Endoplasmic Reticulum Chaperone BiP; D000939:Epitopes; D005434:Flow Cytometry; D006031:Glycosylation; D006360:Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D018832:Molecular Chaperones / Q000378:metabolism*; D009154:Mutation; D010957:Plasmids
chemical_list: D000091342:Endoplasmic Reticulum Chaperone BiP; D000939:Epitopes; C000717247:HSPA5 protein, human; D006360:Heat-Shock Proteins; D018832:Molecular Chaperones