Deglycosylated mammalian beta 2-adrenergic receptors: effect on radioligand binding and peptide mapping.
Mammalian beta-adrenergic receptors are glycoproteins containing both high-mannose and complex-type carbohydrate chains [G. L. Stiles, J. L. Benovic, M. G. Caron, and R. J. Lefkowitz (1984) J. Biol. Chem. 259, 8655-8663]. Endoglycosidase F treatment of beta 2-adrenergic receptors results in the removal of at least two N-linked oligosaccharides, resulting in an increased mobility of the receptor peptide on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (Mr 62,000 to 49,000). In the present study the properties of the deglycosylated beta 2-adrenergic receptor were assessed. Following deglycosylation, the beta 2-adrenergic receptor recognized both agonists and antagonists with the same potency order and affinities as the glycosylated form. In addition, two-dimensional gel electrophoresis peptide mapping techniques applied to control and deglycosylated beta 2-adrenergic receptors (both within and between species) demonstrated that there was a marked homology of the beta 2-adrenergic receptor between species which are closely related phylogenetically. In addition, the glycan component of the receptor did not appear to interfere with the ability of proteinases to generate accurate peptide maps.
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D000818:Animals; D002241:Carbohydrates / Q000302:isolation & purification*; D055598:Chemical Phenomena; D002621:Chemistry; D006224:Cricetinae; D006026:Glycoside Hydrolases; D008168:Lung / Q000378:metabolism; D008297:Male; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase; D010446:Peptide Fragments / Q000032:analysis*; D011869:Radioligand Assay; D051381:Rats; D011919:Rats, Inbred Strains; D011943:Receptors, Adrenergic, beta / Q000378:metabolism*; D012156:Reticulocytes / Q000378:metabolism; D013045:Species Specificity
D002241:Carbohydrates; D010446:Peptide Fragments; D011943:Receptors, Adrenergic, beta; D006026:Glycoside Hydrolases; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase