Dimerization in the absence of higher-order oligomerization of the G protein-coupled secretin receptor

Kaleeckal G. Harikumar, Renee M. Happs, Laurence J. Miller

Research output: Contribution to journalArticle

35 Scopus citations

Abstract

Oligomerization of G protein-coupled receptors has been proposed to affect receptor function and regulation; however, little is known about the molecular nature of such complexes. We previously utilized bioluminescence resonance energy transfer (BRET) to demonstrate that the prototypic Family B secretin receptor can form oligomers. We now explore the order of oligomerization present utilizing unique bimolecular fluorescence complementation and energy transfer techniques. The non-fluorescent carboxyl-terminal and amino-terminal halves of yellow fluorescent protein (YFP) were fused to the carboxyl terminus of the secretin receptor. These constructs bound secretin normally and signaled in response to secretin like wild type receptor. When co-expressed on COS cells, these constructs physically interacted to yield typical YFP fluorescence in biosynthetic compartments and at the plasma membrane, reflecting receptor homo-dimerization. However, the addition of another potential partner in form of Rlu- or CFP-tagged secretin receptor yielded no significant BRET or FRET signal, respectively, under conditions in which intact YFP-tagged secretin receptor yielded such a signal. Absence of higher-order receptor oligomers was further confirmed using saturation BRET techniques. Absence of significant resonance transfer to the secretin receptor homo-dimer was true for carboxyl-terminally-tagged secretin receptor, as well as for receptor incorporating the transfer partner into each of the three distinct intracellular loop domains. These results suggest that the secretin receptor can exist only as a structurally-specific homo-dimer, without being present as higher-order oligomers.

Original languageEnglish (US)
Pages (from-to)2555-2563
Number of pages9
JournalBiochimica et Biophysica Acta - Biomembranes
Volume1778
Issue number11
DOIs
StatePublished - Nov 1 2008

Keywords

  • Bimolecular fluorescence complementation
  • Bioluminescence resonance energy transfer
  • Fluorescence resonance energy transfer
  • G protein-coupled receptor
  • Receptor oligomerization
  • Secretin receptor

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Cell Biology

Fingerprint Dive into the research topics of 'Dimerization in the absence of higher-order oligomerization of the G protein-coupled secretin receptor'. Together they form a unique fingerprint.

  • Cite this