Protein kinase C-mediated inhibition of recombinant T-type CaV3.2 channels by neurokinin 1 receptors
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The voltage-activated T-type calcium channel (Cav3.2) and the G protein-coupled neurokinin 1 (NK1) receptor are expressed in peripheral tissues and in central neurons, in which they participate in diverse physiological processes, including neurogenic inflammation and nociception. In the present report, we demonstrate that recombinant Cav3.2 channels are reversibly inhibited by NK1 receptors when both proteins are transiently coexpressed in human embryonic kidney 293 cells. We found that the voltage-dependent macroscopic properties of Cav3.2 currents were unaffected during NK1 receptor-mediated inhibition. However, inhibition was attenuated in cells coexpressing either the dominant-negative Gαq Q209L/D277N or the regulator of G protein signaling (RGS) proteins 2 (RGS2) and 3T (RGS3T), which are effective antagonists of Gαq/11. By contrast, inhibition was unaffected in cells coexpressing human rod transducin (Gαt), which buffers Gβγ. Channel inhibition was blocked by 1-[6-[[17β-methoxyestra- 1,3,5(10)-trien-17-yl]amino]hexyl]-1Hpyrrole-2,5-dione (U73122) and bisindolylmaleimide I, selective inhibitors of phospholipase Cβ and protein kinase C (PKC), respectively. Inhibition was occluded by application of the PKC activator phorbol-12-myristate-13-acetate. Altogether, these data indicate that NK1 receptors inhibit CaV3.2 channels through a voltage-independent signaling pathway that involves Gαq/11, phospholipase Cβ, and PKC. Our results provide novel evidence regarding the mechanisms underlying T-type calcium channel modulation by G protein-coupled receptors. Functional coupling between Cav3.2 channels and NK1 receptors may be relevant in neurogenic inflammation, neuronal rhythmogenesis, nociception, and other physiological processes. Copyright © 2009 The American Society for Pharmacology and Experimental Therapeutics.
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1 [[6 (3 methoxyestra 1,3,5(10) trien 17beta yl)amino]hexyl] 1h pyrrole 2,5 dione; bisindolylmaleimide; calcium channel T type; G alpha q 11 protein; g alpha q protein; g beta gamma protein; G protein coupled receptor; guanine nucleotide binding protein; guanine nucleotide binding protein alpha subunit; neurokinin 1 receptor; phorbol 13 acetate 12 myristate; phospholipase C beta; protein kinase C; recombinant protein; RGS protein; RGS2 protein; RGS3T protein; transducin; unclassified drug; voltage gated calcium channel; article; attenuation; calcium current; cell strain HEK293; controlled study; embryo; enzyme inhibition; human; human cell; nucleotide sequence; priority journal; protein expression; signal transduction
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