The study of CXCR3 and CCR7 pharmacology using [35S]GTPgammaS exchange assays in cell membranes and permeabilized peripheral blood lymphocytes.
The GTPgammaS exchange assay is a functional model corresponding to the first step of G protein-coupled receptor activation. We provide simple methodologies and controls for setting up GTPgammaS exchange assays in both cell membranes and permeabilized peripheral blood lymphocytes. Specifically, we use guanosine 5'-[35S]triphospate ([35S]GTPgammaS) exchange, in concert with radioligand binding assays, to assess the expression and function of two chemokine receptors important in the trafficking of T lymphocytes: CXCR3 and CCR7. The studies presented here illustrate the utility of GTPgammaS exchange assays in the study of chemokine receptor pharmacology.
Gonsiorek
Waldemar
W
Department of Immunology, Schering-Plough Research Institute, K15 E307C-3945, Kenilworth, NJ 07033-0539, USA.
Zavodny
Paul
P
Hipkin
R William
RW
D001667:Binding, Competitive; D002462:Cell Membrane / Q000378:metabolism; D002463:Cell Membrane Permeability; D016244:Guanosine 5'-O-(3-Thiotriphosphate) / Q000378:metabolism*; D006153:Guanosine Diphosphate / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D008214:Lymphocytes / Q000378:metabolism*; D011869:Radioligand Assay / Q000379:methods; D054400:Receptors, CCR7; D054367:Receptors, CXCR3; D019707:Receptors, Chemokine / Q000378:metabolism*; D012044:Regression Analysis; D012588:Scintillation Counting; D014407:Tumor Cells, Cultured
C515370:CCR7 protein, human; C515187:CXCR3 protein, human; D054400:Receptors, CCR7; D054367:Receptors, CXCR3; D019707:Receptors, Chemokine; D006153:Guanosine Diphosphate; D016244:Guanosine 5'-O-(3-Thiotriphosphate)