Site-Specific Incorporation of Genetically Encoded Photo-Crosslinkers Locates the Heteromeric Interface of a GPCR Complex in Living Cells

Urjita H. Shah, Rudy Toneatti, Supriya A. Gaitonde, Jong M. Shin, Javier González-Maeso

Research output: Contribution to journalArticlepeer-review

16 Scopus citations

Abstract

G protein-coupled receptors (GPCRs) are critical mediators of cell signaling. Although capable of activating G proteins in a monomeric form, numerous studies reveal a possible association of class A GPCRs into dimers/oligomers. The relative location of individual protomers within these GPCR complexes remains a topic of intense debate. We previously reported that class A serotonin 5-HT2A receptor (5-HT2AR) and class C metabotropic glutamate 2 receptor (mGluR2) are able to form a GPCR heterocomplex. By introducing the photoactivatable unnatural amino acid p-azido-L-phenylalanine (azF) at selected individual positions along the transmembrane (TM) segments of mGluR2, we delineate the residues that physically interact at the heteromeric interface of the 5-HT2AR-mGluR2 complex. We show that 5-HT2AR crosslinked with azF incorporated at the intracellular end of mGluR2's TM4, while no crosslinking was observed at other positions along TM1 and TM4. Together, these findings provide important insights into the structural arrangement of the 5-HT2AR-mGluR2 complex.

Original languageEnglish
Pages (from-to)1308-1317.e4
JournalCell Chemical Biology
Volume27
Issue number10
DOIs
StatePublished - 15 Oct 2020
Externally publishedYes

Keywords

  • 5-HT2AR-mGlu2R heteromeric interface
  • GPCRs
  • heterodimers
  • metabotropic glutamate type 2 receptors (mGlu2R)
  • p-azido-L-phenylalanine (azF)
  • photo-crosslinking
  • serotonin 2A receptors (5-HTR)
  • unnatural amino acids

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