Dynamic oligomerization of hRAGE's transmembrane and cytoplasmic domains within SDS micelles

Jamillah Zamoon, Dhanya Madhu, Ikhlas Ahmed

Research output: Contribution to journalArticle

Abstract

The human Receptor for Advanced Glycation End Products (hRAGE) is a pattern recognition receptor implicated in inflammation and adhesion. It is involved in both innate and adaptive immunity. Its aberrant signaling is tied to the pathogenesis of diabetic complications, neurodegenerative disorders, and chronic inflammatory responses. Previous structural studies have focused on its extracellular domains with their canonical constant and variable Ig folds, and to a much lesser extent, the intrinsically disorder cytoplasmic domain. No experimental data are reported on the transmembrane domain, which is integral to signaling. We have constructed, expressed and purified the transmembrane domain attached to the cytoplasmic domain of hRAGE in E. coli. Multiple self-associated forms of these domains were observed in vitro. This pattern of mixed oligomers resembled previously reported in vivo forms of the complete receptor. The self-association of these two domains was further characterized using: SDS-PAGE, intrinsic tryptophan fluorescence and heteronuclear NMR spectroscopy. NMR conditions were assessed across time and temperature within micelles. Our data show that the transmembrane and cytoplasmic domains of hRAGE undergo dynamic oligomerizations that can occur in the absence of its extracellular domains or ligand binding. And, such associations are only partially disrupted even with prolonged incubation in strong detergents.

Original languageEnglish
Pages (from-to)10-18
Number of pages9
JournalInternational Journal of Biological Macromolecules
Volume130
DOIs
Publication statusPublished - 1 Jun 2019

Keywords

  • Amino Acid Sequence
  • Cell Line
  • Cell Membrane/metabolism
  • Cytoplasm/metabolism
  • Humans
  • Micelles
  • Protein Domains/drug effects
  • Protein Multimerization/drug effects
  • Protein Structure, Quaternary/drug effects
  • Receptor for Advanced Glycation End Products/chemistry
  • Sodium Dodecyl Sulfate/chemistry

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