title: Phospholipid mediated activation of calcium dependent protein kinase 1 (CaCDPK1) from chickpea: a new paradigm of regulation.
abstract: Phospholipids, the major structural components of membranes, can also have functions in regulating signaling pathways in plants under biotic and abiotic stress. The effects of adding phospholipids on the activity of stress-induced calcium dependent protein kinase (CaCDPK1) from chickpea are reported here. Both autophosphorylation as well as phosphorylation of the added substrate were enhanced specifically by phosphatidylcholine and to a lesser extent by phosphatidic acid, but not by phosphatidylethanolamine. Diacylgylerol, the neutral lipid known to activate mammalian PKC, stimulated CaCDPK1 but at higher concentrations. Increase in V(max) of the enzyme activity by these phospholipids significantly decreased the K(m) indicating that phospholipids enhance the affinity towards its substrate. In the absence of calcium, addition of phospholipids had no effect on the negligible activity of the enzyme. Intrinsic fluorescence intensity of the CaCDPK1 protein was quenched on adding PA and PC. Higher binding affinity was found with PC (K(½) = 114 nM) compared to PA (K(½) = 335 nM). We also found that the concentration of PA increased in chickpea plants under salt stress. The stimulation by PA and PC suggests regulation of CaCDPK1 by these phospholipids during stress response.
authors.affiliation: Department of Biochemistry, Indian Institute of Science, Bangalore, Karnataka, India.
authors.forename: Ajay Kumar
authors.initials: AK
authors.lastname: Dixit
authors.forename: Chelliah
authors.initials: C
authors.lastname: Jayabaskaran
mesh_terms: D002118:Calcium / Q000378:metabolism; D029863:Cicer; D004075:Diglycerides / Q000494:pharmacology*; D005453:Fluorescence; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects*; D010714:Phosphatidylethanolamines / Q000494:pharmacology*; D010743:Phospholipids / Q000494:pharmacology*; D010766:Phosphorylation / Q000187:drug effects; D011494:Protein Kinases / Q000378:metabolism*; D012965:Sodium Chloride / Q000494:pharmacology; D013312:Stress, Physiological / Q000187:drug effects*; D017871:Calcium-Calmodulin-Dependent Protein Kinases
chemical_list: D002118:Calcium; D004075:Diglycerides; D010714:Phosphatidylethanolamines; D010743:Phospholipids; D011494:Protein Kinases; D012965:Sodium Chloride; D017871:Calcium-Calmodulin-Dependent Protein Kinases; C483858:phosphatidylethanolamine