Functional domains of plant chimeric calcium/calmodulin-dependent protein kinase: regulation by autoinhibitory and visinin-like domains

S Ramachandiran; D Takezawa; W Wang; B W Poovaiah

doi:10.1093/oxfordjournals.jbchem.a021684

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Functional domains of plant chimeric calcium/calmodulin-dependent protein kinase: regulation by autoinhibitory and visinin-like domains

Journal article

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Functional domains of plant chimeric calcium/calmodulin-dependent protein kinase: regulation by autoinhibitory and visinin-like domains

S Ramachandiran, D Takezawa, W Wang and B W Poovaiah

Journal of biochemistry (Tokyo), Vol.121(5), pp.984-990

05/1997

DOI: https://doi.org/10.1093/oxfordjournals.jbchem.a021684

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https://hdl.handle.net/2376/111329

PMID: 9192744

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Abstract

Protein Structure, Tertiary

Amino Acid Sequence

Binding, Competitive - physiology

Molecular Sequence Data

Calcium-Binding Proteins - antagonists & inhibitors

Mutation - genetics

Plants - enzymology

Nerve Tissue Proteins - metabolism

Sequence Homology, Amino Acid

Calcium-Calmodulin-Dependent Protein Kinases - chemistry

Peptides - chemical synthesis

Peptides - metabolism

Plants - genetics

Plant Proteins - chemistry

Gene Deletion

Calmodulin-Binding Proteins - antagonists & inhibitors

Binding, Competitive - genetics

Binding Sites

Calcium-Calmodulin-Dependent Protein Kinases - metabolism

A novel calcium-binding calcium/calmodulin-dependent protein kinase (CCaMK) with a catalytic domain, calmodulin-binding domain, and a neural visinin-like domain was cloned and characterized from plants [Patil et al., (1995) Proc. Natl. Acad. Sci. USA 92, 4797-4801; Takezawa et al. (1996) J. Biol. Chem. 271, 8126-8132]. The mechanisms of CCaMK activation by calcium and calcium/calmodulin were investigated using various deletion mutants. The use of deletion mutants of CCaMK lacking either one, two, or all three calcium-binding EF hands indicated that all three calcium-binding sites in the visinin-like domain were crucial for the full calcium/calmodulin-dependent kinase activity. As each calcium-binding EF hand was deleted, there was a gradual reduction in calcium/calmodulin-dependent kinase activity from 100 to 4%. Another mutant (amino acids 1-322) which lacks both the visinin-like domain containing three EF hands and the calmodulin-binding domain was constitutively active, indicating the presence of an autoinhibitory domain around the calmodulin-binding domain. By using various synthetic peptides and the constitutively active mutant, we have shown that CCaMK contains an autoinhibitory domain within the residues 322-340 which overlaps its calmodulin-binding domain. Kinetic studies with both ATP and the GS peptide substrate suggest that the autoinhibitory domain of CCaMK interacts only with the peptide substrate binding motif of the catalytic domain, but not with the ATP-binding motif.

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Title: Functional domains of plant chimeric calcium/calmodulin-dependent protein kinase: regulation by autoinhibitory and visinin-like domains
Creators: S Ramachandiran - Department of Horticulture, Washington State University, Pullman 99164-6414, USA
D Takezawa
W Wang
B W Poovaiah
Publication Details: Journal of biochemistry (Tokyo), Vol.121(5), pp.984-990
Academic Unit: Department of Horticulture
Publisher: England
Identifiers: 99900547306201842
Language: English
Resource Type: Journal article