Intracellular trypsin effects on the voltage-dependant inactivation of volume-sensitive chloride current in prostate cells
Y.M. Vitko R.N. Lazarenko N.C. Pogorelaya
A.A. Bogomoletz Institute of Physiology NationalAcademy of Sciences of Ukraine, KievРобота підтримана грантом INTAS-99-01248.
Abstract
By means of the patch-clamp technique we have studied
the effects of intracellular applied trypsin, a
known modulator of membrane channel function, on
the properties of the Cl- current induced by hypotonicity-obliged
cell swelling (I Cl, swell) in human
prostate cancer epithelial cells, LNCaP. Intracellular
infusion of 1 mg/ml of trypsin into LNCaP cells via
the patch pipette shortened the delay for the onset
and the time of development of I Cl, swell in response to
hypotonicity as well as accelerated the rate of current
diminution following the return to isotonic conditions.
The maximal density of I Cl, swell in the presence of
intracellular trypsin was 2-fold higher while the current
voltage-dependent inactivation at high depolarizing
potentials was virtually eliminated. Intracellular
co-application of the trypsin inhibitor together
with trypsin abolished all effects of trypsin. We conclude
that VRACs share a great degree of functional
and structural homology to voltage-gated Na+, K+ and
Cl- channels by having intracellular inactivation
domain subjected to proteolytic cleavage that function
in conformity with “ball-and-chain” inactivation model.
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