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Updated: Aug 16, 2026

Pull-down of Calmodulin-binding Proteins
Published on: January 23, 2012
Tween-20 activates and solubilizes the mitochondrial membrane-bound, calmodulin dependent NAD+ finase of Avena sativa
M A Pou De Crescenzo1, S Gallais, A Léon
1Equipe de Biochimie et de Biologie Moléculaire, UFR Sciences, Université d'Angers, 2 Boulevard Lavoisier, 49 045 Angers Cedex 01, France.
Abstract:
Among different treatments assayed, a mix of a nonionic detergent (5% Tween-20) with 0.5 m NaCl was found to solubilize a large part of the calmodulin-dependent NAD+ kinase bound to the inner mitochondrial membrane. It also stimulated its activity by increasing 7 times the maximal velocity. Activity stimulation was also observed with phosphatidylcholine, phosphatidylethanolamine and with reductants (HSO3 and DTT). This solubilized NAD+ kinase and the calmodulin-dependent cytosoluble isoform displayed distinct molecular masses, as well as different kinetic parameters. We propose that solubilization of membrane-bound NAD+ kinase could occur in vivo in Avena sativa and could generate a soluble isoform.
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