Characterization of the swine adipocyte A1 adenosine receptor using an optimized assay system.
The radioligand binding assay of A1 adenosine receptors in adipocyte crude plasma membrane from Yucatan miniature swine was optimized by evaluating 17 factors involved in the assay. Significant effects of CHAPS, adenosine deaminase, EDTA, pre-rinsing glass fiber filters and pH were found for the binding measurements. Using the optimized procedure, [3H]8-cyclopentyl-1,3-dipropylxanthine, ([3H]-DPCPX) binding to A1 adenosine receptors in swine subcutaneous adipocyte crude plasma membrane was measured; Bmax and Kd values were 479 +/- 77 fmol/mg protein and 0.87 +/- 0.10 nM, respectively. Values for mesenteric adipose tissue from sedentary swine and subcutaneous adipose tissue from exercise-trained swine were also measured.
Dong
Q
Q
Department of Animal and Nutritional Sciences, University of New Hampshire, Durham 03824.
Schuchman
J
J
Carey
G B
GB
D000243:Adenosine Deaminase / Q000378:metabolism; D017667:Adipocytes / Q000378:metabolism*; D000818:Animals; D001665:Binding Sites; D002462:Cell Membrane / Q000201:enzymology; D002793:Cholic Acids; D005260:Female; D006863:Hydrogen-Ion Concentration; D008274:Magnesium / Q000494:pharmacology; D011480:Protease Inhibitors / Q000494:pharmacology; D058915:Purinergic P1 Receptor Antagonists; D018047:Receptors, Purinergic P1 / Q000378:metabolism*; D013552:Swine; D013556:Swine, Miniature; D014970:Xanthines / Q000494:pharmacology
D002793:Cholic Acids; D011480:Protease Inhibitors; D058915:Purinergic P1 Receptor Antagonists; D018047:Receptors, Purinergic P1; D014970:Xanthines; C051360:1,3-dipropyl-8-cyclopentylxanthine; D000243:Adenosine Deaminase; D008274:Magnesium; C028213:3-((3-cholamidopropyl)dimethylammonium)-1-propanesulfonate