Negative regulation of ligand-initiated Ca2+uptake by PKC-βII in differentiated HL60 cells

Abstract

In phagocytic cells, fMet-Leu-Phe triggers phosphoinositide remodeling, activation of protein kinase C (PKC), release of intracellular Ca(2+) and uptake of extracellular Ca(2+). Uptake of extracellular Ca(2+) can be triggered by store-operated Ca(2+) channels (SOCC) and via a receptor-operated nonselective cation channel(s). In neutrophilic HL60 cells, the PKC activator phorbol myristate acetate (PMA) activates multiple PKC isotypes, PKC-alpha, PKC-beta, and PKC-delta, and inhibits ligand-initiated mobilization of intracellular Ca(2+) and uptake of extracellular Ca(2+). Therefore PKC is a negative regulator at several points in Ca(2+) mobilization. In contrast, selective depletion of PKC-beta in HL60 cells by an antisense strategy enhanced fMet-Leu-Phe-initiated Ca(2+) uptake but not mobilization of intracellular Ca(2+). Thapsigargin-induced Ca(2+) uptake through SOCC was not affected by PKC-beta II depletion. Thus PKC-beta II is a selective negative regulator of Ca(2+) uptake but not release of intracellular Ca(2+) stores. PKC-beta II inhibits a receptor-operated cation or Ca(2+) channel, thus inhibiting ligand-initiated Ca(2+) uptake.