title: Purified rat hepatic beta 2-adrenergic receptor. Structural similarities to the rat fat cell beta 1-adrenergic receptor.
abstract: The mammalian beta 2-adrenergic receptor from rat liver has been purified by sequential cycles of affinity chromatography followed by steric exclusion high performance liquid chromatography. In purified preparations, the overall yield of receptor approaches 10%. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of highly purified receptor preparations reveals a single peptide, Mr = 67,000, as judged by silver staining. Purified beta 2-adrenergic receptor migrates on steric-exclusion high performance liquid chromatography in two peaks, Mr = 140,000 and 67,000. Specific binding of (-)-[3H]dihydroalprenolol and (-)-[125I]iodocyanopindolol to purified rat liver beta-adrenergic receptor preparations is stereoselective and displays a rank order of potencies characteristic of a beta 2-adrenergic receptor. The mammalian beta 1-adrenergic receptor of rat fat cells has also been purified (Cubero, A., and Malbon, C.C. (1984) J. Biol. Chem. 259, 1344-1350). When purified in the presence of protease inhibitors, radioiodinated beta 1-adrenergic receptors from rat fat cells and beta 2-adrenergic receptors from rat liver comigrate on sodium dodecyl sulfate-polyacrylamide gels as 67,000 Mr peptides. Autoradiograms of two-dimensional partial proteolytic digests of the purified, radioiodinated rat liver beta-adrenergic receptor, as generated by alpha-chymotrypsin, Staphylococcus aureus V8 protease, and elastase reveal a pattern of peptide fragments essentially identical to those generated by partial proteolytic digests of the purified radioiodinated beta 1-receptor from rat fat cells. This data suggests that a high degree of homology exists between these two pharmacologically distinct mammalian beta-adrenergic receptor proteins.
authors.forename: M P
authors.initials: MP
authors.lastname: Graziano
authors.forename: C P
authors.initials: CP
authors.lastname: Moxham
authors.forename: C C
authors.initials: CC
authors.lastname: Malbon
mesh_terms: D000273:Adipose Tissue / Q000378:metabolism*; D000818:Animals; D001667:Binding, Competitive; D002462:Cell Membrane / Q000378:metabolism; D002846:Chromatography, Affinity / Q000379:methods; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004072:Digitonin; D004082:Dihydroalprenolol / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D020120:Iodocyanopindolol; D007700:Kinetics; D008099:Liver / Q000378:metabolism*; D008970:Molecular Weight; D009928:Organ Specificity; D010869:Pindolol / Q000031:analogs & derivatives / Q000378:metabolism; D051381:Rats; D011919:Rats, Inbred Strains; D011943:Receptors, Adrenergic, beta / Q000302:isolation & purification* / Q000378:metabolism
chemical_list: D011943:Receptors, Adrenergic, beta; D004082:Dihydroalprenolol; D020120:Iodocyanopindolol; D010869:Pindolol; D004072:Digitonin