title: Disulfide bonds within the C2 domain of RAGE play key roles in its dimerization and biogenesis.
abstract: BACKGROUND
  The receptor for advanced glycation end products (RAGE) on the cell surface transmits inflammatory signals. A member of the immunoglobulin superfamily, RAGE possesses the V, C1, and C2 ectodomains that collectively constitute the receptor's extracellular structure. However, the molecular mechanism of RAGE biogenesis remains unclear, impeding efforts to control RAGE signaling through cellular regulation.  METHODOLOGY AND RESULT: We used co-immunoprecipitation and crossing-linking to study RAGE oligomerization and found that RAGE forms dimer-based oligomers. Via non-reducing SDS-polyacrylamide gel electrophoresis and mutagenesis, we found that cysteines 259 and 301 within the C2 domain form intermolecular disulfide bonds. Using a modified tripartite split GFP complementation strategy and confocal microscopy, we also found that RAGE dimerization occurs in the endoplasmic reticulum (ER), and that RAGE mutant molecules without the double disulfide bridges are unstable, and are subjected to the ER-associated degradation.
  
  
  CONCLUSIONS
  Disulfide bond-mediated RAGE dimerization in the ER is the critical step of RAGE biogenesis. Without formation of intermolecular disulfide bonds in the C2 region, RAGE fails to reach cell surface.
  
  
  CONCLUSIONS
  This is the first report of RAGE intermolecular disulfide bond.
authors.affiliation: Laboratory of Cardiovascular Science, National Institute on Aging, Baltimore, Maryland, United States of America.
authors.forename: Wen
authors.initials: W
authors.lastname: Wei
authors.forename: Leonie
authors.initials: L
authors.lastname: Lampe
authors.forename: Sungha
authors.initials: S
authors.lastname: Park
authors.forename: Bhavana S
authors.initials: BS
authors.lastname: Vangara
authors.forename: Geoffrey S
authors.initials: GS
authors.lastname: Waldo
authors.forename: Stephanie
authors.initials: S
authors.lastname: Cabantous
authors.forename: Sarah S
authors.initials: SS
authors.lastname: Subaran
authors.forename: Dongmei
authors.initials: D
authors.lastname: Yang
authors.forename: Edward G
authors.initials: EG
authors.lastname: Lakatta
authors.forename: Li
authors.initials: L
authors.lastname: Lin
mesh_terms: D000818:Animals; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D003545:Cysteine / Q000737:chemistry; D004220:Disulfides / Q000737:chemistry*; D004721:Endoplasmic Reticulum / Q000378:metabolism; D005786:Gene Expression Regulation; D055786:Gene Knockout Techniques; D006031:Glycosylation; D006367:HeLa Cells; D006801:Humans; D051379:Mice; D055503:Protein Multimerization; D020836:Protein Structure, Quaternary; D017434:Protein Structure, Tertiary; D000067759:Receptor for Advanced Glycation End Products; D011971:Receptors, Immunologic / Q000096:biosynthesis* / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction
chemical_list: D004220:Disulfides; D000067759:Receptor for Advanced Glycation End Products; D011971:Receptors, Immunologic; D003545:Cysteine