Characterization of constitutively active mutants of G protein-coupled receptors.
The ability of G protein-coupled receptors to transduce signaling typically is induced by the binding of an appropriate ligand (agonist), resulting in a conformational change of the receptor and the subsequent interaction with the G protein heterotrimer. Some mutants of G protein-coupled receptors, known as constitutively active mutants, have the capacity to activate the G protein-signaling cascade even in the absence of ligand. In this chapter, we describe three methods that most directly allow characterization of constitutively active mutants and discriminate them from the wild-type receptors. All methods are based on the spontaneous signaling function in the absence of ligand and its consequences on the receptor.
Navenot
Jean-Marc
JM
Department of Pathology, Medical College of Georgia, Augusta, USA.
Wang
Zi-xuan
ZX
Peiper
Stephen C
SC
D000818:Animals; D016466:CHO Cells; D006224:Cricetinae; D016244:Guanosine 5'-O-(3-Thiotriphosphate) / Q000378:metabolism; D006801:Humans; D008024:Ligands; D009154:Mutation; D010766:Phosphorylation; D011485:Protein Binding; D019718:Receptors, CXCR4 / Q000235:genetics / Q000378:metabolism; D043562:Receptors, G-Protein-Coupled / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015290:Second Messenger Systems / Q000502:physiology*
D008024:Ligands; D019718:Receptors, CXCR4; D043562:Receptors, G-Protein-Coupled; D016244:Guanosine 5'-O-(3-Thiotriphosphate)