Proteolytic modification of swelling-activated Cl- current in LNCaP prostate cancer epithelial cells.

The effects of intracellular application of trypsin on the Cl- current induced by hypotonic cell swelling (I(Cl,swell)) in human prostate cancer epithelial cells (LNCaP) was studied using the patch-clamp technique. In cells predialyzed with 1 mg/mL trypsin, I(Cl,swell)) developed and diminished in response to the application and withdrawal of hypotonic solution about three times faster than that in control cells. In trypsin-infused cells, I(Cl,swell)) also had about twofold higher current density and displayed considerably slowed voltage-dependent inactivation, which was quite pronounced in control cells at potentials above +60 mV. Trypsin-induced modification of I(Cl,swell)) could be prevented by coinfusion of 10 mg/mL soybean trypsin inhibitor, suggesting that proteolytic cleavage of essential intracellular structural domains of the I(Cl,swell))-carrying volume-regulated anion channel (VRAC) was responsible for this functional modification. The effect of trypsin was not dependent on the presence of intracellular ATP. We conclude that VRACs, similarly to voltage-gated Na+, K+, and Cl- channels, possess intracellular inactivation domain(s) subjected to proteolytic cleavage that may function in conformity with the classical "ball-and-chain" inactivation model.