Analysis of G-protein-coupled receptor kinase RGS homology domains.
G-protein-coupled receptor kinases (GRKs) specifically phosphorylate agonist-occupied G-protein-coupled receptors (GPCRs). All seven mammalian GRKs contain an N-terminal domain that is homologous to the regulator of G-protein signaling (RGS) family of proteins. The RGS domain of GRK2 has been shown to interact specifically with Galphaq family members. While the specificity and functional consequences of GRK/Galpha interaction remain somewhat poorly defined, GRK RGS homology (RH) domains likely function to provide specificity for GRK interaction with a particular Galpha subunit or GPCR/Galpha complex. Indeed, GRK2 binds Galphaq, alpha11, and alpha14, but not Galpha16, Galphas, Galphai, or Galpha(12/13), while the RGS domains of GRK5 and GRK6 do not bind Galpha(q/11), Galphas, Galphai, or Galpha(12/13). In this chapter we describe various in vitro and intact cell strategies that can be used to elucidate the function of GRK RH domains.
Day
Peter W
PW
Department of Microbiology and Immunology, Thomas Jefferson University, Philadelphia, Pennsylvania 19107, USA.
Wedegaertner
Philip B
PB
Benovic
Jeffrey L
JL
D000818:Animals; D001681:Biological Assay / Q000379:methods; D002460:Cell Line; D044385:GTP-Binding Protein alpha Subunits / Q000378:metabolism; D019204:GTP-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D007295:Inositol Phosphates / Q000378:metabolism; D011485:Protein Binding; D017346:Protein Serine-Threonine Kinases / Q000235:genetics / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D021122:Protein Subunits / Q000378:metabolism; D020710:RGS Proteins / Q000378:metabolism*; D043562:Receptors, G-Protein-Coupled / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D015290:Second Messenger Systems / Q000502:physiology*
D044385:GTP-Binding Protein alpha Subunits; D007295:Inositol Phosphates; D021122:Protein Subunits; D020710:RGS Proteins; D043562:Receptors, G-Protein-Coupled; D011993:Recombinant Fusion Proteins; D017346:Protein Serine-Threonine Kinases; D019204:GTP-Binding Proteins