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.
Department of Microbiology and Immunology, Thomas Jefferson University, Philadelphia, Pennsylvania 19107, USA.
Peter W
PW
Day
Philip B
PB
Wedegaertner
Jeffrey L
JL
Benovic
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