PURIFICATION AND FUNCTIONAL-CHARACTERIZATION OF THE HUMAN-BETA-2-ADRENERGIC RECEPTOR PRODUCED IN BACULOVIRUS-INFECTED INSECT CELLS

被引:54
作者
REILANDER, H
BOEGE, F
VASUDEVAN, S
MAUL, G
HEKMAN, M
DEES, C
HAMPE, W
HELMREICH, EJM
MICHEL, H
机构
[1] UNIV WURZBURG,MED POLIKLIN,W-8700 WURZBURG,GERMANY
[2] UNIV WURZBURG,INST PHYSIOL CHEM,W-8700 WURZBURG,GERMANY
关键词
BETA-ADRENERGIC RECEPTOR; BACULOVIRUS; EXPRESSION; GLYCOSYLATION; AFFINITY CHROMATOGRAPHY;
D O I
10.1016/0014-5793(91)80532-8
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
A human cDNA fragment bearing the complete coding region for the beta2-adrenergic receptor was introduced into the genome of Autographa california nuclear polyhedrosis virus under the control of the polyhedrin promoter. Binding studies using [I-125]iodocyanopindolol showed that Sf9 insect cells infected with the recombinant virus expressed almost-equal-to 1 x 10(6) beta-2-adrenergic receptors on their cell surface. Photoaffinity labeling of whole cells and membranes revealed a molecular weight of almost-equal-to 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.
引用
收藏
页码:441 / 444
页数:4
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