BIOCHEMICAL-CHARACTERIZATION AND SOLUBILIZATION OF HUMAN NK2 RECEPTOR EXPRESSED IN CHINESE-HAMSTER OVARY CELLS

被引:11
作者
TURCATTI, G [1 ]
CESZKOWSKI, K [1 ]
CHOLLET, A [1 ]
机构
[1] GLAXO INST MOLEC BIOL SA,14 CHEMIN AULX,CH-1228 PLAN OUATES GENEV,SWITZERLAND
来源
JOURNAL OF RECEPTOR RESEARCH | 1993年 / 13卷 / 1-4期
关键词
D O I
10.3109/10799899309073684
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
The human ileum neurokinin NK2 receptor has been stably expressed in Chinese hamster ovary (CHO) cells using the dihydrofolate reductase (DHFR) expression system. Amplified cell populations expressing approximately 7x10(5) NK2 receptors/cell were selected in the presence of the DHFR inhibitor methotrexate. Cross-linking of [I-125]NKA to NK2 receptor transfected cells revealed a specifically labeled protein of apparent molecular weight 64 kDa by SDS-polyacrylamide gel electrophoresis. This protein was deglycosylated by the enzymes N-glycosidase F and endoglycosydase F to a protein of apparent molecular weight of 39 kDa. The NK2 receptor was solubilized in an active form from CHO cell membranes using the zwitterionic detergent CHAPS. This method represents a valuable approach for the production of significant amounts of NK2 receptor protein from mammalian cells.
引用
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页码:639 / 652
页数:14
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