FUNCTIONAL EXPRESSION OF THE RAT ANION-EXCHANGER AE2 IN INSECT CELLS BY A RECOMBINANT BACULOVIRUS

被引:26
|
作者
HE, XJ
WU, XZ
KNAUF, PA
TABAK, LA
MELVIN, JE
机构
[1] UNIV ROCHESTER, DEPT DENT RES, BOX 611, 601 ELMWOOD AVE, ROCHESTER, NY 14642 USA
[2] UNIV ROCHESTER, DEPT NEUROBIOL & ANAT, ROCHESTER, NY 14642 USA
[3] UNIV ROCHESTER, DEPT BIOCHEM, ROCHESTER, NY 14642 USA
[4] UNIV ROCHESTER, DEPT BIOPHYS, ROCHESTER, NY 14642 USA
来源
AMERICAN JOURNAL OF PHYSIOLOGY | 1993年 / 264卷 / 04期
关键词
SF9; CELLS; COMPLEMENTARY DEOXYRIBONUCLEIC ACID EXPRESSION; 4,4'-DIISOTHIOCYANOSTILBENE-2,2'-DISULFONATE; 2'; 7'-BIS(2-CARBOXYETHYL)-5(6)-CARBOXYFLUORESCEIN; INTRACELLULAR PH;
D O I
10.1152/ajpcell.1993.264.4.C1075
中图分类号
Q4 [生理学];
学科分类号
071003 ;
摘要
We used baculovirus to transiently express a rat anion exchanger (AE2) in Spodoptera frugiperda (Sf9) insect cells. No detectable Cl--HCO3- exchange activity was observed in wild type or sham-infected Sf9 cells, monitored using 2',7'-bis(2 - carboxyethyl) - 5(6) - carboxyfluorescein, a pH-sensitive fluorescent dye. Functional expression of anion exchange activity in the AE2 recombinant baculovirus-infected cells was observed within the first day after infection and sustained over the next 3 days. The expressed AE2 anion exchange activity was Na+ independent and could be reversibly and irreversibly inhibited by the specific anion exchange inhibitor 4,4-diisothiocyanostilbene-2,2'-disulfonate (DIDS). The reversible inhibition was sensitive to the concentration of DIDS, with a half inhibition of 4 muM. These results indicate that the rat AE2 protein produced in the recombinant baculovirus-infected insect cells is inserted into the plasma membrane in a biologically active form that appears suitable for functional studies of AE2.
引用
收藏
页码:C1075 / C1079
页数:5
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