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Am J Physiol Cell Physiol 264: C1075-C1079, 1993;
0363-6143/93 $5.00
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AJP - Cell Physiology, Vol 264, Issue 4 C1075-C1079, Copyright © 1993 by American Physiological Society

ARTICLES

Functional expression of the rat anion exchanger AE2 in insect cells by a recombinant baculovirus

X. He, X. Wu, P. A. Knauf, L. A. Tabak and J. E. Melvin
Department of Biochemistry, University of Rochester, New York 14642.

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 microM. 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.


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