Actinin-1 binds to the C-terminus of A2B adenosine receptor (A2BAR) and enhances A2BAR cell-surface expression

Biochem J. 2016 Jul 15;473(14):2179-86. doi: 10.1042/BCJ20160272. Epub 2016 May 17.

Abstract

A2BAR (A2B adenosine receptor) has been implicated in several physiological conditions, such as allergic or inflammatory disorders, vasodilation, cell growth and epithelial electrolyte secretion. For mediating the protein-protein interactions of A2BAR, the receptor's C-terminus is recognized to be crucial. In the present study, we unexpectedly found that two point mutations in the A2BAR C-terminus (F297A and R298A) drastically impaired the expression of A2BAR protein by accelerating its degradation. Thus we tested the hypothesis that these two point mutations disrupt A2BAR's interaction with a protein essential for A2BAR stability. Our results show that both mutations disrupted the interaction of A2BAR with actinin-1, an actin-associated protein. Furthermore, actinin-1 binding stabilized the global and cell-surface expression of A2BAR. By contrast, actinin-4, another non-muscle actinin isoform, did not bind to A2BAR. Thus our findings reveal a previously unidentified regulatory mechanism of A2BAR abundance.

Keywords: A2B adenosine receptor; G-protein-coupled receptor (GPCR); actinin; stabilization; surface expression.

MeSH terms

  • Actinin / metabolism*
  • Animals
  • COS Cells
  • Chlorocebus aethiops
  • Cyclic AMP / metabolism
  • HEK293 Cells
  • Humans
  • Immunoprecipitation
  • Point Mutation / genetics
  • Protein Binding / genetics
  • Protein Binding / physiology
  • Receptor, Adenosine A2B / chemistry
  • Receptor, Adenosine A2B / genetics
  • Receptor, Adenosine A2B / metabolism*
  • Signal Transduction

Substances

  • Receptor, Adenosine A2B
  • Actinin
  • Cyclic AMP