A fully active nonglycosylated V2 vasopressin receptor.
The human V2 vasopressin receptor belongs to the superfamily of G protein-coupled receptors believed to be anchored to the plasma membrane by seven transmembrane regions. The extracellular portion of the human V2 vasopressin receptor contains one site susceptible to N-linked glycosylation. Metabolic labeling and immunoprecipitation of the receptor expressed in transfected cells were applied to examine whether the protein was indeed glycosylated. The V2 vasopressin receptor expressed transiently was glycosylated, but glycosidase treatment to test the complexity of the sugar moiety linked to asparagine revealed that the majority of the receptor protein lacked complex carbohydrates, an indication of an improperly processed protein. This immature protein displayed a tendency to form aggregates. In contrast with these data, testing of the sugar complexity of the receptor protein synthesized in stably transfected cells identified the predominant form as an appropriately processed receptor protein. Mutagenesis of asparagine 22 to glutamine produced on expression in transfected cells a nonglycosylated receptor with ligand binding affinity and coupling characteristics almost identical to those of the wild-type form. After exposure to elevated concentrations of AVP (100 nM), the nonglycosylated form desensitized to the same extent as the wild-type receptor.
Innamorati
G
G
Department of Anesthesiology and Molecular Biology Institute, UCLA School of Medicine 90095, USA.
Sadeghi
H
H
Birnbaumer
M
M
D000262:Adenylyl Cyclases / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D001127:Arginine Vasopressin / Q000378:metabolism / Q000494:pharmacology*; D002460:Cell Line; D002522:Chlorocebus aethiops; D000939:Epitopes; D019204:GTP-Binding Proteins / Q000378:metabolism; D015870:Gene Expression; D006026:Glycoside Hydrolases; D006031:Glycosylation; D006801:Humans; D007668:Kidney; D007700:Kinetics; D008969:Molecular Sequence Data; D008970:Molecular Weight; D016297:Mutagenesis, Site-Directed; D016133:Polymerase Chain Reaction; D017483:Receptors, Vasopressin / Q000096:biosynthesis* / Q000737:chemistry / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification; D014162:Transfection
D000939:Epitopes; D017483:Receptors, Vasopressin; D011994:Recombinant Proteins; D001127:Arginine Vasopressin; D006026:Glycoside Hydrolases; D019204:GTP-Binding Proteins; D000262:Adenylyl Cyclases