title: Regulation of Receptor for Advanced Glycation End Products (RAGE) Ectodomain Shedding and Its Role in Cell Function.
abstract: The receptor for advanced glycation end products (RAGE) is a multiligand transmembrane receptor that can undergo proteolysis at the cell surface to release a soluble ectodomain. Here we observed that ectodomain shedding of RAGE is critical for its role in regulating signaling and cellular function. Ectodomain shedding of both human and mouse RAGE was dependent on ADAM10 activity and induced with chemical activators of shedding (ionomycin, phorbol 12-myristate 13-acetate, and 4-aminophenylmercuric acetate) and endogenous stimuli (serum and RAGE ligands). Ectopic expression of the splice variant of RAGE (RAGE splice variant 4), which is resistant to ectodomain shedding, inhibited RAGE ligand dependent cell signaling, actin cytoskeleton reorganization, cell spreading, and cell migration. We found that blockade of RAGE ligand signaling with soluble RAGE or inhibitors of MAPK or PI3K blocked RAGE-dependent cell migration but did not affect RAGE splice variant 4 cell migration. We finally demonstrated that RAGE function is dependent on secretase activity as ADAM10 and γ-secretase inhibitors blocked RAGE ligand-mediated cell migration. Together, our data suggest that proteolysis of RAGE is critical to mediate signaling and cell function and may therefore emerge as a novel therapeutic target for RAGE-dependent disease states.
authors.affiliation: From the Department of Cell Biology and Department of Biochemistry, Leonard M. Miller School of Medicine, University of Miami, Miami, Florida 33136.
authors.forename: Alex
authors.initials: A
authors.lastname: Braley
authors.affiliation: From the Department of Cell Biology and Department of Biochemistry, Leonard M. Miller School of Medicine, University of Miami, Miami, Florida 33136.
authors.forename: Taekyoung
authors.initials: T
authors.lastname: Kwak
authors.affiliation: From the Department of Cell Biology and Department of Biochemistry, Leonard M. Miller School of Medicine, University of Miami, Miami, Florida 33136.
authors.forename: Joel
authors.initials: J
authors.lastname: Jules
authors.affiliation: From the Department of Cell Biology and Department of Biochemistry, Leonard M. Miller School of Medicine, University of Miami, Miami, Florida 33136.
authors.forename: Evis
authors.initials: E
authors.lastname: Harja
authors.affiliation: From the Department of Cell Biology and Department of Biochemistry, Leonard M. Miller School of Medicine, University of Miami, Miami, Florida 33136.
authors.forename: Ralf
authors.initials: R
authors.lastname: Landgraf
authors.affiliation: From the Department of Cell Biology and Department of Biochemistry, Leonard M. Miller School of Medicine, University of Miami, Miami, Florida 33136 bhudson@med.miami.edu.
authors.forename: Barry I
authors.initials: BI
authors.lastname: Hudson
mesh_terms: D000072197:ADAM10 Protein / Q000378:metabolism; D000595:Amino Acid Sequence; D053829:Amyloid Precursor Protein Secretases / Q000378:metabolism; D000818:Animals; D001665:Binding Sites / Q000235:genetics; D015153:Blotting, Western; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000187:drug effects / Q000235:genetics / Q000502:physiology*; D002468:Cell Physiological Phenomena / Q000187:drug effects / Q000235:genetics / Q000502:physiology*; D057809:HEK293 Cells; D006801:Humans; D015759:Ionomycin / Q000494:pharmacology; D045726:Metalloproteases / Q000378:metabolism; D051379:Mice; D009154:Mutation; D020033:Protein Isoforms / Q000235:genetics / Q000378:metabolism; D059748:Proteolysis / Q000187:drug effects; D000067759:Receptor for Advanced Glycation End Products / Q000235:genetics / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid; D015398:Signal Transduction / Q000187:drug effects / Q000235:genetics / Q000502:physiology*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
chemical_list: D020033:Protein Isoforms; D000067759:Receptor for Advanced Glycation End Products; D015759:Ionomycin; D053829:Amyloid Precursor Protein Secretases; D045726:Metalloproteases; D000072197:ADAM10 Protein; D013755:Tetradecanoylphorbol Acetate
keywords: ADAM; S100 proteins; cell migration; cell signaling; ectodomain shedding; matrix metalloproteinase (MMP); receptor for advanced glycation end products (RAGE)