Antibodies against a furosemide binding protein from Ehrlich ascites tumour cells inhibit Na+, K+, Cl- co-transport in frog retinal pigment epithelium.

Abstract

Purpose: To investigate whether antibodies against a 100 kDa protein purified by furosemide affinity chromatography from Ehrlich ascites tumour cells could inhibit Na+, K+, Cl- co-transport in the isolated frog retinal pigment epithelium.

Methods: The rate of Na+, K+, Cl- co-transport across the retinal membrane in the isolated frog RPE preparation was measured as the rate of decrease in the intracellular Cl- activity observed after administration of furosemide in the apical bath. The intracellular Cl- activity was measured with double barrelled Cl- sensitive microelectrodes.

Results: Incubation of frog retinal pigment epithelium for 30 min with antibodies reduced the rate of Na+, K+, Cl- co-transport by 43%, while leaving all other measured electrophysiological parameters intact.

Conclusion: The antibodies inhibit Na+,K+,Cl- co-transport in the frog retinal pigment epithelium. This could be due to binding of the antibodies to the co-transporter itself or to a regulatory protein.