Reconstitution of G protein-coupled receptors with recombinant G protein alpha and beta gamma subunits

Methods Enzymol. 2002:343:372-93. doi: 10.1016/s0076-6879(02)43146-1.

Abstract

The methods outlined in this article describe experiments that can probe the first steps in receptor:G protein interaction using defined, recombinant receptors and G proteins. The protocols have the advantages that the receptors are inserted properly in a cell membrane and that the investigator has complete control of the proteins reconstituted with the receptor. Specific mutations in the receptors or G proteins are studied easily and the protocols allow precise examination of the stoichiometry of the receptor:alpha:beta gamma interaction.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Baculoviridae / genetics
  • Cells, Cultured
  • GTP-Binding Proteins / chemistry
  • GTP-Binding Proteins / metabolism*
  • RNA, Antisense
  • Receptors, Cell Surface / genetics
  • Receptors, Cell Surface / metabolism*
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / metabolism
  • Sensitivity and Specificity
  • Transfection
  • Urea / chemistry

Substances

  • RNA, Antisense
  • Receptors, Cell Surface
  • Recombinant Proteins
  • Urea
  • GTP-Binding Proteins