The fat cell beta-adrenergic receptor. Purification and characterization of a mammalian beta 1-adrenergic receptor.
The beta 1-adrenergic receptor of rat fat cells was effectively solubilized with digitonin and purified by affinity chromatography and steric exclusion high pressure liquid chromatography (HPLC). The purification strategy described permits an approximately 24,000-fold purification of the beta 1-adrenergic receptor of fat cells with an overall recovery of approximately 70%. Purified receptor preparations demonstrate a specific activity for (-) [3H]dihydroalprenolol binding of 12 nmol/mg of protein. The purified receptor was shown to migrate in steric exclusion HPLC as a Mr = 67,000 protein. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of radioiodinated purified receptor revealed a single, major peptide of Mr = 67,000. The binding of (-) [3H]dihydroalprenolol to purified receptor preparations displayed stereoselectivity and affinities for antagonists similar in nature to the membrane-bound and digitonin-solubilized beta 1-adrenergic receptor. In addition to the Mr = 67,000 component, a Mr = 140,000 form of the receptor was identified in HPLC runs of freshly prepared, affinity chromatographed receptor preparations that had not been frozen. This larger form of the receptor yielded binding activity of Mr = 67,000 on sequential HPLC runs and was shown to contain the Mr = 67,000 peptide. The beta 1-receptor from this mammalian source, composed of a single Mr = 67,000 peptide, is clearly quite distinct from the purified avian beta 1-, amphibian beta 2-, and mammalian beta 2-adrenergic receptors described by others.
D000273:Adipose Tissue / Q000032:analysis*; D000818:Animals; D002846:Chromatography, Affinity; D002851:Chromatography, High Pressure Liquid; D004072:Digitonin; D004082:Dihydroalprenolol / Q000378:metabolism; D005260:Female; D008970:Molecular Weight; D051381:Rats; D011919:Rats, Inbred Strains; D011943:Receptors, Adrenergic, beta / Q000302:isolation & purification*; D012995:Solubility
D011943:Receptors, Adrenergic, beta; D004082:Dihydroalprenolol; D004072:Digitonin