Mutational analysis of beta-adrenergic receptor glycosylation.
The beta-adrenergic receptor (beta AR) contains significant amounts of N-linked carbohydrate. Deletion mutants spanning the four consensus glycosylation sites on the receptor and single amino acid substitutions within the two amino-terminal consensus glycosylation sites reveal that both the amino-terminal sites are utilized. None of the glycosylation-defective beta AR mutants exhibited altered ligand binding in transient expression assays. In addition, the mutant beta ARs which were completely devoid of carbohydrate were capable of coupling to Gs and stimulating adenylyl cyclase in stable L cell lines. In contrast to the wild-type beta AR, the glycosylation-deficient beta ARs expressed in these cells showed a 50% decrease in the level of accumulation on the cell surface. Therefore, while glycosylation of the beta AR does not appear to be essential for receptor function, it is important for correct trafficking of the beta AR protein through the cell.
Rands
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E
Department of Molecular Biology, Merck, Sharp and Dohme Research Laboratories, West Point, Pennsylvania 19486.
Candelore
M R
MR
Cheung
A H
AH
Hill
W S
WS
Strader
C D
CD
Dixon
R A
RA
D000262:Adenylyl Cyclases / Q000378:metabolism; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D002872:Chromosome Deletion; D003001:Cloning, Molecular; D006224:Cricetinae; D004247:DNA / Q000235:genetics; D005944:Glucosamine / Q000378:metabolism; D006031:Glycosylation; D006165:Guanylyl Imidodiphosphate / Q000494:pharmacology; D007545:Isoproterenol / Q000494:pharmacology; D008969:Molecular Sequence Data; D009154:Mutation; D015345:Oligonucleotide Probes; D011943:Receptors, Adrenergic, beta / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D014162:Transfection; D014415:Tunicamycin / Q000494:pharmacology
D015345:Oligonucleotide Probes; D011943:Receptors, Adrenergic, beta; D014415:Tunicamycin; D006165:Guanylyl Imidodiphosphate; D004247:DNA; D000262:Adenylyl Cyclases; D007545:Isoproterenol; D005944:Glucosamine