title: Mapping the Interactome of a Major Mammalian Endoplasmic Reticulum Heat Shock Protein 90.
abstract: Up to 10% of cytosolic proteins are dependent on the mammalian heat shock protein 90 (HSP90) for folding. However, the interactors of its endoplasmic reticulum (ER) paralogue (gp96, Grp94 and HSP90b1) has not been systematically identified. By combining genetic and biochemical approaches, we have comprehensively mapped the interactome of gp96 in macrophages and B cells. A total of 511 proteins were reduced in gp96 knockdown cells, compared to levels observed in wild type cells. By immunoprecipitation, we found that 201 proteins associated with gp96. Gene Ontology analysis indicated that these proteins are involved in metabolism, transport, translation, protein folding, development, localization, response to stress and cellular component biogenesis. While known gp96 clients such as integrins, Toll-like receptors (TLRs) and Wnt co-receptor LRP6, were confirmed, cell surface HSP receptor CD91, TLR4 pathway protein CD180, WDR1, GANAB and CAPZB were identified as potentially novel substrates of gp96. Taken together, our study establishes gp96 as a critical chaperone to integrate innate immunity, Wnt signaling and organ development.
authors.affiliation: Department of Microbiology & Immunology, Medical University of South Carolina, Charleston, United States of America.
authors.forename: Feng
authors.initials: F
authors.lastname: Hong
authors.affiliation: Department of Microbiology & Immunology, Medical University of South Carolina, Charleston, United States of America.
authors.forename: Saleh
authors.initials: S
authors.lastname: Mohammad Rachidi
authors.affiliation: Center for Vascular Biology, University of Connecticut Health Center, Farmington, Connecticut, United States of America.
authors.forename: Debbie
authors.initials: D
authors.lastname: Lundgren
authors.affiliation: Center for Vascular Biology, University of Connecticut Health Center, Farmington, Connecticut, United States of America.
authors.forename: David
authors.initials: D
authors.lastname: Han
authors.affiliation: Department of Epidemiology and Public Health, Yale University, School of Medicine, New Haven, Connecticut, United States of America.
authors.forename: Xiu
authors.initials: X
authors.lastname: Huang
authors.affiliation: Department of Epidemiology and Public Health, Yale University, School of Medicine, New Haven, Connecticut, United States of America.
authors.forename: Hongyu
authors.initials: H
authors.lastname: Zhao
authors.affiliation: Laboratory for Immunogenomics, RIKEN Research Center for Allergy and Immunology, Tsurumi-ku, Yokohama, Kanagawa, Japan.
authors.forename: Yayoi
authors.initials: Y
authors.lastname: Kimura
authors.affiliation: Laboratory for Immunogenomics, RIKEN Research Center for Allergy and Immunology, Tsurumi-ku, Yokohama, Kanagawa, Japan.
authors.forename: Hisashi
authors.initials: H
authors.lastname: Hirano
authors.affiliation: Graduate School of Nanobioscience, Yokohama City University, Tsurumi-ku, Yokohama, Kanagawa, Japan.|Kazusa DNA Research Institute, Kisarazu, Chiba, Japan.
authors.forename: Osamu
authors.initials: O
authors.lastname: Ohara
authors.affiliation: Department of Immunology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Japan.
authors.forename: Heichiiro
authors.initials: H
authors.lastname: Udono
authors.affiliation: CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences (CAS), Beijing, China.
authors.forename: Songdong
authors.initials: S
authors.lastname: Meng
authors.affiliation: Department of Microbiology & Immunology, Medical University of South Carolina, Charleston, United States of America.
authors.forename: Bei
authors.initials: B
authors.lastname: Liu
authors.affiliation: Department of Microbiology & Immunology, Medical University of South Carolina, Charleston, United States of America.
authors.forename: Zihai
authors.initials: Z
authors.lastname: Li
mesh_terms: D000818:Animals; D001402:B-Lymphocytes / Q000378:metabolism; D015153:Blotting, Western; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D002853:Chromatography, Liquid; D015180:Electrophoresis, Gel, Two-Dimensional; D004721:Endoplasmic Reticulum / Q000378:metabolism*; D005838:Genotype; D018841:HSP90 Heat-Shock Proteins / Q000378:metabolism*; D047468:Immunoprecipitation; D008562:Membrane Glycoproteins / Q000378:metabolism; D051379:Mice; D018832:Molecular Chaperones / Q000378:metabolism; D015398:Signal Transduction / Q000502:physiology; D053719:Tandem Mass Spectrometry
chemical_list: D018841:HSP90 Heat-Shock Proteins; D008562:Membrane Glycoproteins; D018832:Molecular Chaperones