A protein kinase activity associated with maize root plasma membranes was partially purified and characterized. Biochemical properties, such as calcium dependence, inhibition by calmodulin antagonists, and absence of calmodulin stimulation; indicated that the enzyme belongs to the calcium-dependent protein kinase (CDPK) family. By means of an in-gel phosphorylation assay the molecular mass of active polypeptides was determined: two bands of 55 and 51 kDa became labelled. The same proteins were also immunodecorated by monoclonal antibodies against soybean CDPK. Results from in vitro assays demonstrated that maize H+-ATPase was a suitable substrate for this protein kinase and that the phosphorylation site was located at the C-terminal domain of the enzyme. This result was confirmed by using as substrate in phosphorylation assays the isolated C-terminal domain of the H+- ATPase expressed in Escherichia coli as a glutathione-transferase fusion protein.

The plasma membrane H+-ATPase from maize roots is phosphorylated in the C-terminal domain by a calcium-dependent protein kinase / L., Camoni; Fullone, Maria Rosaria; M., Marra; P., Aducci. - In: PHYSIOLOGIA PLANTARUM. - ISSN 0031-9317. - STAMPA. - 104(1998), pp. 549-555.

The plasma membrane H+-ATPase from maize roots is phosphorylated in the C-terminal domain by a calcium-dependent protein kinase

FULLONE, Maria Rosaria;
1998

Abstract

A protein kinase activity associated with maize root plasma membranes was partially purified and characterized. Biochemical properties, such as calcium dependence, inhibition by calmodulin antagonists, and absence of calmodulin stimulation; indicated that the enzyme belongs to the calcium-dependent protein kinase (CDPK) family. By means of an in-gel phosphorylation assay the molecular mass of active polypeptides was determined: two bands of 55 and 51 kDa became labelled. The same proteins were also immunodecorated by monoclonal antibodies against soybean CDPK. Results from in vitro assays demonstrated that maize H+-ATPase was a suitable substrate for this protein kinase and that the phosphorylation site was located at the C-terminal domain of the enzyme. This result was confirmed by using as substrate in phosphorylation assays the isolated C-terminal domain of the H+- ATPase expressed in Escherichia coli as a glutathione-transferase fusion protein.
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Utilizza questo identificativo per citare o creare un link a questo documento: http://hdl.handle.net/11573/123671
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