Purification and functional characterization of the human beta 2-adrenergic receptor produced in baculovirus-infected insect cells.
A human cDNA fragment bearing the complete coding region for the beta 2-adrenergic receptor was introduced into the genome of Autographa california nuclear polyhedrosis virus under the control of the polyhedrin promoter. Binding studies using [125I]iodocyanopindolol showed that Sf9 insect cells infected with the recombinant virus expressed approximately 1 x 10(6) beta 2-adrenergic receptors on their cell surface. Photoaffinity labeling of whole cells and membranes revealed a molecular weight of approximately 46,000 for the expressed receptor. The receptor produced in insect cells is glycosylated but the extent and pattern differ from that of the receptor from human tissue. The heterologously expressed receptor was purified by alprenolol affinity chromatography, and was able to activate isolated Gs-protein.
Reiländer
H
H
Max Planck Institut für Biophysik, Frankfurt, Germany.
Boege
F
F
Vasudevan
S
S
Maul
G
G
Hekman
M
M
Dees
C
C
Hampe
W
W
Helmreich
E J
EJ
Michel
H
H
D000262:Adenylyl Cyclases / Q000378:metabolism; D000526:Alprenolol / Q000378:metabolism; D016367:Baculoviridae; D003001:Cloning, Molecular; D019204:GTP-Binding Proteins / Q000378:metabolism; D005822:Genetic Vectors; D006031:Glycosylation; D006801:Humans; D066298:In Vitro Techniques; D046911:Macromolecular Substances; D011943:Receptors, Adrenergic, beta / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D011994:Recombinant Proteins / Q000302:isolation & purification*
D046911:Macromolecular Substances; D011943:Receptors, Adrenergic, beta; D011994:Recombinant Proteins; D000526:Alprenolol; D019204:GTP-Binding Proteins; D000262:Adenylyl Cyclases