Disulfide Bonds within the C2 Domain of RAGE Play Key Roles in Its Dimerization and Biogenesis

Wen Wei, Leonie Lampe, Sungha Park, Bhavana S. Vangara, Geoffrey S. Waldo, Stephanie Cabantous, Sarah S. Subaran, Dongmei Yang, Edward G. Lakatta, Li Lin

Research output: Contribution to journalArticle

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. Conclusion: 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. Significance: This is the first report of RAGE intermolecular disulfide bond.

Original languageEnglish (US)
Article numbere50736
JournalPloS one
Volume7
Issue number12
DOIs
StatePublished - Dec 17 2012
Externally publishedYes

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)
  • General

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    Wei, W., Lampe, L., Park, S., Vangara, B. S., Waldo, G. S., Cabantous, S., Subaran, S. S., Yang, D., Lakatta, E. G., & Lin, L. (2012). Disulfide Bonds within the C2 Domain of RAGE Play Key Roles in Its Dimerization and Biogenesis. PloS one, 7(12), [e50736]. https://doi.org/10.1371/journal.pone.0050736