Cell surface expression of the human N-methyl-D-aspartate receptor subunit 1a requires the co-expression of the NR2A subunit in transfected cells

被引:84
作者
McIlhinney, RAJ [1 ]
Molnar, E [1 ]
Atack, JR [1 ]
Whiting, PJ [1 ]
机构
[1] MERCK SHARP & DOHME RES LABS,NEUROSCI RES CTR,HARLOW CM20 2QR,ESSEX,ENGLAND
关键词
NMDA receptors; transfection; ion channels; plasma membrane; biotinylation; antibody;
D O I
10.1016/0306-4522(95)00419-X
中图分类号
Q189 [神经科学];
学科分类号
071006 ;
摘要
Cell surface expression of the NR1a subunit has been examined in mouse L cell lines permanently transfected with the complementary DNA for human NR1a or with the complementary DNAs for NR1a and NR2A. The expression of the subunits was under the control of the murine mammary tumour virus promoter and following induction of expression by dexamethazone both cell lines expressed high levels of the NR1a subunit as determined by immunofluorescence using permeabilized cells and immunoblotting of cell membranes with subunit specific antibodies. However, cell surface expression of the NR1a subunit was found only in the cells expressing both the NR1a and NR2A subunits. This was confirmed by cell surface biotinylation of the two cell lines and affinity isolation of the receptor subunits. To determine if this result was solely due to the use of a particular cell line and or the choice of expression vector, Cos-7 cells were transiently transfected with either NR1a or NR1a plus NR2A. Here too cell surface expression was only found following co-transfection of both subunits.
引用
收藏
页码:989 / 997
页数:9
相关论文
共 46 条
[1]   HOMOMERIC AND NATIVE ALPHA-7 ACETYLCHOLINE-RECEPTORS EXHIBIT REMARKABLY SIMILAR BUT NONIDENTICAL PHARMACOLOGICAL PROPERTIES, SUGGESTING THAT THE NATIVE RECEPTOR IS A HETEROMERIC PROTEIN COMPLEX [J].
ANAND, R ;
PENG, X ;
LINDSTROM, J .
FEBS LETTERS, 1993, 327 (02) :241-246
[2]   COMBINATORIAL RNA SPLICING ALTERS THE SURFACE-CHARGE ON THE NMDA RECEPTOR [J].
ANANTHARAM, V ;
PANCHAL, RG ;
WILSON, A ;
KOLCHINE, VV ;
TREISTMAN, SN ;
BAYLEY, H .
FEBS LETTERS, 1992, 305 (01) :27-30
[3]  
ANEGAWA NJ, 1995, J NEUROCHEM, V64, P2004
[4]   STABLE TRANSFECTION OF THE NR1 SUBUNIT IN CHINESE-HAMSTER OVARY CELLS FAILS TO PRODUCE A FUNCTIONAL N-METHYL-D-ASPARTATE RECEPTOR [J].
BOECKMAN, FA ;
AIZENMAN, E .
NEUROSCIENCE LETTERS, 1994, 173 (1-2) :189-192
[5]  
BROSE N, 1993, J BIOL CHEM, V268, P22663
[6]   IMMUNOLOGICAL DETECTION OF THE NMDAR1 GLUTAMATE RECEPTOR SUBUNIT EXPRESSED IN HUMAN EMBRYONIC KIDNEY-293 CELLS AND IN RAT-BRAIN [J].
CHAZOT, PL ;
CIK, M ;
STEPHENSON, FA .
JOURNAL OF NEUROCHEMISTRY, 1992, 59 (03) :1176-1178
[7]  
CHAZOT PL, 1994, J BIOL CHEM, V269, P24403
[8]   OPTIMAL EXPRESSION OF CLONED NMDAR1 NMDAR2A HETEROMERIC GLUTAMATE RECEPTORS - A BIOCHEMICAL-CHARACTERIZATION [J].
CIK, M ;
CHAZOT, PL ;
STEPHENSON, FA .
BIOCHEMICAL JOURNAL, 1993, 296 :877-883
[9]   FIBROBLASTS TRANSFECTED WITH TORPEDO ACETYLCHOLINE-RECEPTOR BETA-SUBUNIT, GAMMA-SUBUNIT, AND DELTA-SUBUNIT CDNAS EXPRESS FUNCTIONAL RECEPTORS WHEN INFECTED WITH A RETROVIRAL ALPHA-RECOMBINANT [J].
CLAUDIO, T ;
PAULSON, HL ;
GREEN, WN ;
ROSS, AF ;
HARTMAN, DS ;
HAYDEN, D .
JOURNAL OF CELL BIOLOGY, 1989, 108 (06) :2277-2290
[10]  
CULLEN BR, 1987, METHOD ENZYMOL, V152, P684