| 7559508 |
Schumaker KS, Gizinski MJ: 1,4-Dihydropyridine binding sites in moss plasma membranes. J Biol Chem. 1995 Oct 6;270(40):23461-7.
Properties of receptors for a calcium channel antagonist.. An increase in cytoplasmic calcium is an early event in hormone (cytokinin)-induced vegetative bud formation in the moss Physcomitrella patens. Whole cell and calcium transport studies have implicated 1,4-dihydropyridine-sensitive calcium channels in this increase in cellular calcium. To understand the molecular nature of the dihydropyridine-sensitive calcium channel, we have established conditions for the binding of the arylazide 1,4-dihydropyridine, [3H] azidopine, to its receptor in moss plasma membranes. [3H] Azidopine bound specifically in a saturable and reversible manner. The KD for [3H] azidopine binding was 5.2 nM and the Bmax was 35.6 pmol/mg of protein. Association and dissociation of the receptor and [3H] azidopine were temperature-dependent, and association varied as a function of pH. Binding was inhibited by dihydropyridine, phenylalkylamine, and benzothiazepine calcium channel blockers, bepridil, lanthanum, and N-ethylmaleimide. [3H] Azidopine binding was stimulated by cations including calcium, strontium, manganese, and barium. [3H] Azidopine binding was also stimulated by cytokinin with a Km value for kinetin of 0.13 nM. These studies utilize a simple plant system to provide a biochemical framework for understanding calcium regulation during development and have implications for understanding mechanisms of signal transduction in plants. |
33(0,1,1,3) |