Cytosolic Ca2+ and protein kinase Cα couple cellular metabolism to membrane K+ permeability in a human biliary cell line

Yu Wang, Richard Roman, Thorsten Schlenker, Yusuf A. Hannun, John Raymond, J. Gregory Fitz

Research output: Contribution to journalArticlepeer-review

25 Scopus citations

Abstract

Cholangiocytes represent an important target of injury during the ischemia and metabolic stress that accompanies liver preservation. Since K+ efflux serves to minimize injury during ATP depletion in certain other cell types, the purpose of these studies was to evaluate the effects of ATP depletion on plasma membrane K+ permeability of Mz-ChA-1 cells, a model human biliary cell line. Cells were exposed to dinitrophenol (50 μM) and 2- deoxyglucose (10 mM) as the standard model of metabolic injury. Whole-cell and single K+ channel currents were measured using patch clamp techniques; and intracellular [Ca2+] ([Ca2+](i)) was estimated by calcium green-1 fluorescence. Metabolic stress increased [Ca2+](i), and stimulated translocation of the α isoform of protein kinase C (PKCα) from cytosolic to particulate cell fractions. The same maneuver increased membrane K+ permeability 40-70-fold as detected by (a) activation of K+-selective whole cell currents of 2,176±218 pA (n = 34), and (b) opening of apamin-sensitive K+ channels with a unitary conductance of 17.0±0.2 pS. PKCα translocation and channel opening appear to be related since stress-induced K+ efflux is inhibited by chelation of cytosolic Ca2+, exposure to the PKC inhibitor chelerythrine (25 μM) and downregulation of PKC by phorbol esters. Moreover, K+ currents were activated by intracellular perfusion with recombinant PKCα in the absence of metabolic inhibitors. These findings indicate that in biliary cells apamin-sensitive K+ channels are functionally coupled to cell metabolism and suggest that cytosolic Ca2+ and PKCα are selectively involved in the response.

Original languageEnglish (US)
Pages (from-to)2890-2897
Number of pages8
JournalJournal of Clinical Investigation
Volume99
Issue number12
DOIs
StatePublished - Jun 15 1997

Keywords

  • ATP
  • Cholangiocyte
  • Ion channel
  • Liver

ASJC Scopus subject areas

  • Medicine(all)

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