Regulation of the electrogenic H+ channel in the plasma membrane of neutrophils: possible role of phospholipase A2, internal and external protons.
Possible factors regulating the opening of and the rate of H+ flux through a recently described, Cd(2+)-sensitive, phorbol ester- and arachidonic acid (AA)-activatable H(+)-conducting pathway in the plasma membrane of neutrophil granulocytes were investigated. (1) The phospholipase A2 blocker p-bromophenacyl bromide (BPB) inhibited the phorbol 12-myristate 13-acetate (PMA)-induced activation of this channel in a concentration-dependent manner (IC50, 4 microM). (2) Neither BPB nor the protein kinase C (PKC) inhibitor staurosporine influenced the AA-elicited stimulation of this route. (3) Intracellular acidification (cytoplasmic pH below 6.9) itself is capable of activating an electrogenic, Cd(2+)-sensitive H+ efflux indicating that protons can open up this route in the absence of any other stimulator. (4) PMA significantly decreases the intracellular H+ concentration ([H+]i) threshold for the opening of the channel, thus providing a conductive state at resting pH values, and elevates the rate of H+ efflux at any [H+]i. (5) Changes in external pH also modify the operation of the channel: above an extracellular pH (pH(o)) value of 7.4, the H(+)-flux/driving force relationship is approx. 5-fold greater than below this value. Our results suggest a multifactorial regulation of the electrogenic H+ channel: most probably PKC activates the channel indirectly, via stimulation of phospholipase A2 that subsequently liberates AA. In addition to this, the channel conductance seems to be promoted by internal H+ and inhibited by external H+.
Kapus
A
A
Department of Physiology, Semmelweis Medical University, Budapest, Hungary.
Suszták
K
K
Ligeti
E
E
D000818:Animals; D002104:Cadmium / Q000378:metabolism; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism / Q000502:physiology; D002478:Cells, Cultured; D003593:Cytoplasm / Q000378:metabolism; D004789:Enzyme Activation; D006859:Hydrogen / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D015640:Ion Channel Gating; D007473:Ion Channels / Q000378:metabolism*; D008564:Membrane Potentials; D009504:Neutrophils / Q000187:drug effects / Q000378:metabolism* / Q000502:physiology; D010741:Phospholipases A / Q000502:physiology*; D054467:Phospholipases A2; D011493:Protein Kinase C / Q000378:metabolism; D013552:Swine; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
D007473:Ion Channels; D002104:Cadmium; D006859:Hydrogen; D011493:Protein Kinase C; D010741:Phospholipases A; D054467:Phospholipases A2; D013755:Tetradecanoylphorbol Acetate