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. 1992 Mar;98(3):1196–1201. doi: 10.1104/pp.98.3.1196

Plasma Membrane Ca-ATPase of Radish Seedlings 1

I. Biochemical Characterization Using ITP as a Substrate

Antonella Carnelli 1,2, Maria I De Michelis 1,2, Franca Rasi-Caldogno 1,2
PMCID: PMC1080327  PMID: 16668746

Abstract

In this work, we exploited the capability of the plasma membrane Ca-ATPase to utilize ITP as a substrate to study its characteristics in plasma membrane vesicles purified from radish (Raphanus sativus L.) seedlings. The majority of the ITPase activity of plasma membrane was Ca2+-dependent. The Ca2+-dependent ITPase activity was Mg2+-dependent and was stimulated by the calcium ionophore A23187. It was inhibited by erythrosin B (concentration giving 50% inhibition, 50 nanomolar) and by vanadate (concentration giving 50% inhibition, 3 micromolar) and displayed a broad pH optimum around pH 7.2 to 7.5. Both the hydrolytic and the transport activity of the plasma membrane Ca-ATPase were half-saturated by Ca2+ in the micromolar concentration range. No major effect of EGTA on the saturation kinetics of the enzyme was observed. The affinity of the plasma membrane Ca-ATPase for Ca2+ was about fourfold higher at pH 7.5 than at pH 6.9. The Ca2+-dependent ITPase activity was stimulated about twofold by polyoxyethylene 20 cetyl ether, although it was inhibited by Triton X-100 and by lysolecithin.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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