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Plant and Cell Physiology, 1985, Vol. 26, No. 1 201-209
© 1985

Article

Identification of Naturally Occurring Calmodulin Inhibitors in Plants and their Effects on Calcium- and Calmodulin-promoted Protein Phosphorylation1,2

Gopinadhan Paliyath and B. W. Poovaiah3

Department of Horticulture and Landscape Architecture, Washington State University Pullman, WA 99164-6414, U.S.A.

3 To whom requests for reprints should be addressed

While studying the calmodulin activity in post-climacteric apples, a heat stable, dialyzable component that inhibited calmodulin-promoted phosphodiesterase activity was detected. The compound(s) that inhibited calmodulin activity did not bind to Dowex-50, H+ or Dowex-2, Cl– and was exclusively present in the neutral fraction. The inhibitors irreversibly bound to polyvinylpolypyrrolidone indicating their phenolic nature. Fractionation of the neutral fraction on a C18-µbondapak column and analysis for the inhibition of calmodulin-promoted phosphodiesterase activity showed significant inhibitory activity associated with fractions eluted 5 min, 15 min and 18 min after injection. Perdeuteriomethylation and combined gas chromatographic-mass spectrometric analysis of the inhibitors showed them to be flavonoids. (+)-Catechin was identified in the fraction eluted 5 min after injection that also showed maximum inhibition. Other flavonoids such as epicatechin, quercetin and naringenin also inhibited calmodulin-promoted phosphodiesterase activity. Among the phenolic compounds commonly encountered in plant tissue only caffeic acid inhibited calmodulin-promoted phosphodiesterase activity. Inhibition by catechin nd caffeic acid could be reversed by increasing the calmodulin concentration in the assay mixture. Both catechin and caffeic acid inhibited Ca- and calmodulin-promoted phosphorylation of soluble proteins from corn coleoptiles. The physiological properties of flavonoids are discussed in light of this evidence.

1 Scientific Paper No. 6965, College of Agriculture and Home Economics Research Center, Washington State University, Pullman, Project 0321.

2 Supported in part by grants from the Washington State Tree Fruit Research Commission, American Cancer Society # IN119D and National Science Foundation PCM-8208408.

(Received July 30, 1984; Accepted November 12, 1984)
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