FUNCTIONAL-ACTIVITY OF A BIOTINYLATED HUMAN NEUROKININ-1 RECEPTOR FUSION EXPRESSED IN THE SEMLIKI-FOREST-VIRUS SYSTEM

被引:11
作者
LUNDSTROM, K
VARGAS, A
ALLET, B
机构
[1] Department of Molecular Biology, Glaxo Institute for Molecular Biology, Geneva
关键词
D O I
10.1006/bbrc.1995.1332
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
The 1.3 S biotinylatable subunit of Proprionibacterium shermanii transcarboxylase complex was fused to the C-terminus of the human neurokinin 1 receptor gene and introduced into the Semliki Forest virus expression vector pSFV1. RNA transcribed from pSFV1-NK1-biot and pSFV-Helper2 was coelectroporated into BHK cells permitting in vivo packaging of recombinant virus. Infection of BHK and CHO cells with SFV-NK1-biot virus yielded high level of the fusion receptor as detected by metabolic labeling, immunoblotting with streptavidin alkaline phosphatase and binding to substance P. Like native receptor, the biotinylated receptor fusion was able to stimulate Ca2+ mobilization in infected CHO cells,indicating functional coupling to guanine-nucleotide-binding proteins. (C) 1995 Academic Press, Inc.
引用
收藏
页码:260 / 266
页数:7
相关论文
共 24 条
[1]   MODULATION OF NEUROGENIC INFLAMMATION - NOVEL APPROACHES TO INFLAMMATORY DISEASE [J].
BARNES, PJ ;
BELVISI, MG ;
ROGERS, DF .
TRENDS IN PHARMACOLOGICAL SCIENCES, 1990, 11 (05) :185-189
[2]   SEMLIKI FOREST VIRUS EXPRESSION SYSTEM - PRODUCTION OF CONDITIONALLY INFECTIOUS RECOMBINANT PARTICLES [J].
BERGLUND, P ;
SJOBERG, M ;
GAROFF, H ;
ATKINS, GJ ;
SHEAHAN, BJ ;
LILJESTROM, P .
BIO-TECHNOLOGY, 1993, 11 (08) :916-920
[3]  
CRONAN JE, 1990, J BIOL CHEM, V265, P10327
[4]   VECTORS THAT FACILITATE THE EXPRESSION AND PURIFICATION OF FOREIGN PEPTIDES IN ESCHERICHIA-COLI BY FUSION TO MALTOSE-BINDING PROTEIN [J].
DIGUAN, C ;
LI, P ;
RIGGS, PD ;
INOUYE, H .
GENE, 1988, 67 (01) :21-30
[5]   RECOMBINANT BOVINE NEUROKININ-2 RECEPTOR STABLY EXPRESSED IN CHINESE-HAMSTER OVARY CELLS COUPLES TO MULTIPLE SIGNAL TRANSDUCTION PATHWAYS [J].
EISTETTER, HR ;
CHURCH, DJ ;
MILLS, A ;
GODFREY, PP ;
CAPPONI, AM ;
BREWSTER, R ;
SCHULZ, MF ;
KAWASHIMA, E ;
ARKINSTALL, SJ .
CELL REGULATION, 1991, 2 (10) :767-779
[6]   THE EXTRACELLULAR DOMAIN OF THE NEUROKININ-1 RECEPTOR IS REQUIRED FOR HIGH-AFFINITY BINDING OF PEPTIDES [J].
FONG, TM ;
YU, H ;
HUANG, RRC ;
STRADER, CD .
BIOCHEMISTRY, 1992, 31 (47) :11806-11811
[7]  
FONG TM, 1992, MOL PHARMACOL, V41, P24
[8]  
FONG TM, 1993, NATURE, P350
[9]   EXPRESSION OF 2 HUMAN BETA-ADRENERGIC RECEPTORS IN ESCHERICHIA-COLI - FUNCTIONAL INTERACTION WITH 2 FORMS OF THE STIMULATORY G-PROTEIN [J].
FREISSMUTH, M ;
SELZER, E ;
MARULLO, S ;
SCHUTZ, W ;
STROSBERG, AD .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1991, 88 (19) :8548-8552
[10]   DIFFERENT BINDING EPITOPES ON THE NK1 RECEPTOR FOR SUBSTANCE-P AND A NONPEPTIDE ANTAGONIST [J].
GETHER, U ;
JOHANSEN, TE ;
SNIDER, RM ;
LOWE, JA ;
NAKANISHI, S ;
SCHWARTZ, TW .
NATURE, 1993, 362 (6418) :345-348