Linked Life Data

11399751

Source:http://linkedlifedata.com/resource/pubmed/id/11399751

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
35
pubmed:dateCreated
2001-8-27
pubmed:abstractText
The existence of two molecular switches regulating plant chimeric Ca(2+)/calmodulin-dependent protein kinase (CCaMK), namely the C-terminal visinin-like domain acting as Ca(2+)-sensitive molecular switch and calmodulin binding domain acting as Ca(2+)-stimulated autophosphorylation-sensitive molecular switch, has been described (Sathyanarayanan, P. V., Cremo, C. R., and Poovaiah, B. W. (2000) J. Biol. Chem. 275, 30417-30422). Here we report the identification of Ca(2+)-stimulated autophosphorylation site of CCaMK by matrix-assisted laser desorption ionization time of flight-mass spectrometry. Thr(267) was confirmed as the Ca(2+)-stimulated autophosphorylation site by post-source decay experiments and by site-directed mutagenesis. The purified T267A mutant form of CCaMK did not show Ca(2+)-stimulated autophosphorylation, autophosphorylation-dependent variable calmodulin affinity, or Ca(2+)/calmodulin stimulation of kinase activity. Sequence comparison of CCaMK from monocotyledonous plant (lily) and dicotyledonous plant (tobacco) suggests that the autophosphorylation site is conserved. This is the first identification of a phosphorylation site specifically responding to activation by second messenger system (Ca(2+) messenger system) in plants. Homology modeling of the kinase and calmodulin binding domain of CCaMK with the crystal structure of calcium/calmodulin-dependent protein kinase 1 suggests that the Ca(2+)-stimulated autophosphorylation site is located on the surface of the kinase and far from the catalytic site. Analysis of Ca(2+)-stimulated autophosphorylation with increasing concentration of CCaMK indicates the possibility that the Ca(2+)-stimulated phosphorylation occurs by an intermolecular mechanism.
pubmed:keyword
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Aug
pubmed:issn
0021-9258
pubmed:author
pubmed:issnType
Print
pubmed:day
31
pubmed:volume
276
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
32940-7
pubmed:dateRevised
2007-11-15
pubmed:meshHeading
pubmed-meshheading:11399751-Amino Acid Sequence, pubmed-meshheading:11399751-Amino Acid Substitution, pubmed-meshheading:11399751-Binding Sites, pubmed-meshheading:11399751-Calcium, pubmed-meshheading:11399751-Calcium-Calmodulin-Dependent Protein Kinases, pubmed-meshheading:11399751-Chromatography, Affinity, pubmed-meshheading:11399751-Kinetics, pubmed-meshheading:11399751-Liliaceae, pubmed-meshheading:11399751-Models, Molecular, pubmed-meshheading:11399751-Molecular Sequence Data, pubmed-meshheading:11399751-Mutagenesis, Site-Directed, pubmed-meshheading:11399751-Peptide Fragments, pubmed-meshheading:11399751-Phosphopeptides, pubmed-meshheading:11399751-Phosphorylation, pubmed-meshheading:11399751-Protein Conformation, pubmed-meshheading:11399751-Protein Structure, Secondary, pubmed-meshheading:11399751-Recombinant Fusion Proteins, pubmed-meshheading:11399751-Spectrometry, Mass, Matrix-Assisted Laser...
pubmed:year
2001
pubmed:articleTitle
Calcium-stimulated autophosphorylation site of plant chimeric calcium/calmodulin-dependent protein kinase.
pubmed:affiliation
Laboratory of Plant Molecular Biology and Physiology, Department of Horticulture,Washington State University, Pullman, Washington 99164, USA.
pubmed:publicationType
Journal Article, Research Support, U.S. Gov't, Non-P.H.S.