A novel protein kinase from Brassica juncea stimulated by a protozoan calcium binding protein. Purification and partial characterization.
A novel protein kinase (BjCCaBPk) from etiolated Brassica juncea seedlings has been purified and partially characterized. The purified enzyme migrated on SDS/PAGE as a single band with an apparent molecular mass of 43 kDa. The optimum pH for the kinase activity was 8.0. It was stimulated more than sixfold by the protozoa Entamoeba histolytica calcium binding protein EhCaBP (10.5 nM) but not by calmodulin (CaM) when used at equimolar concentration. Moreover the kinase also did not bind CaM-Sepharose. There was neither inhibition of the kinase activity in the presence of W-7 (a CaM antagonist), KN-62 (a specific calcium/CaM kinase inhibitor) and anti-CaM Ig, nor any effect on BjCCaBPk activity of staurosporine (a protein kinase C inhibitor). Furthermore a CaM-kinase specific substrate, syntide-2, proved to be a poor substrate for the BjCCaBPk compared with histone III-S. The phosphorylation of histone III-S involved serine residues. Southern and Northern blot analysis showed the presence of EhCaBP homologues in Brassica. The data suggest that BjCCaBPk may be a novel protein kinase with an affinity towards a calcium binding protein like EhCaBP.
Deswal
R
R
National Center for Plant Genome Research, Jawaharlal Nehru University Campus, New Delhi, India.
Pandey
G K
GK
Chandok
M R
MR
Yadav
N
N
Bhattacharya
A
A
Sopory
S K
SK
D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine / Q000031:analogs & derivatives / Q000494:pharmacology; D000818:Animals; D001937:Brassica / Q000201:enzymology*; D002118:Calcium / Q000494:pharmacology; D020013:Calcium Signaling; D002135:Calcium-Binding Proteins / Q000494:pharmacology*; D002147:Calmodulin / Q000037:antagonists & inhibitors / Q000494:pharmacology; D002614:Chelating Agents / Q000494:pharmacology; D004533:Egtazic Acid / Q000494:pharmacology; D004748:Entamoeba histolytica / Q000378:metabolism; D006657:Histones / Q000378:metabolism; D007074:Immunoglobulin G / Q000494:pharmacology; D036341:Intercellular Signaling Peptides and Proteins; D010455:Peptides / Q000378:metabolism; D010766:Phosphorylation; D010768:Phosphoserine / Q000032:analysis; D010940:Plant Proteins / Q000302:isolation & purification* / Q000378:metabolism; D011494:Protein Kinases / Q000302:isolation & purification* / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D015800:Protozoan Proteins / Q000494:pharmacology*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D019311:Staurosporine / Q000494:pharmacology; D013449:Sulfonamides / Q000494:pharmacology
D002135:Calcium-Binding Proteins; D002147:Calmodulin; D002614:Chelating Agents; D006657:Histones; D007074:Immunoglobulin G; D036341:Intercellular Signaling Peptides and Proteins; D010455:Peptides; D010940:Plant Proteins; D015800:Protozoan Proteins; D011993:Recombinant Fusion Proteins; D013449:Sulfonamides; C063391:syntide-2; C075554:calcium-binding protein, Entamoeba histolytica; D010768:Phosphoserine; D004533:Egtazic Acid; C063302:KN 62; C017967:W 7; D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; D011494:Protein Kinases; D019311:Staurosporine; D002118:Calcium