Protein kinase C in Trypanosoma cruzi epimastigote forms: partial purification and characterization.
A protein kinase C activity from epimastigote forms of Trypanosoma cruzi was characterized. Cytosolic extracts were chromatographed on DEAE-cellulose columns giving two peaks of kinase activity which were eluted at 0.1 and 0.15 M NaCl. The first activity peak requires Ca2+ and phosphatidylserine for activity. Further kinase purification was performed by chromatography on phenyl Sepharose columns. In these columns the enzyme activity was adsorbed in the presence of Ca2+ and eluted with a EGTA-containing buffer. T. cruzi protein kinase C activity preferentially phosphorylated histone H1. It was stimulated by diacylglycerol and phorbol myristate acetate, and inhibited by polymyxin B and staurosporine. After subcellular fractionation and epimastigote cells, the kinase was found to be associated with microsomal and cytosolic fractions.
Gómez
M L
ML
Instituto de Investigaciones en Ingeniería Genética y Biología Molecular (INGEBI), Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Argentina.
Erijman
L
L
Arauzo
S
S
Torres
H N
HN
Téllez-Iñón
M T
MT
D000818:Animals; D002118:Calcium / Q000494:pharmacology; D002847:Chromatography, Agarose; D002848:Chromatography, DEAE-Cellulose; D003600:Cytosol / Q000201:enzymology; D004075:Diglycerides / Q000494:pharmacology; D004533:Egtazic Acid / Q000494:pharmacology; D006657:Histones / Q000378:metabolism; D008861:Microsomes / Q000201:enzymology; D010718:Phosphatidylserines / Q000494:pharmacology; D010766:Phosphorylation; D011493:Protein Kinase C / Q000302:isolation & purification / Q000378:metabolism*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014349:Trypanosoma cruzi / Q000201:enzymology*
D004075:Diglycerides; D006657:Histones; D010718:Phosphatidylserines; D004533:Egtazic Acid; D011493:Protein Kinase C; D013755:Tetradecanoylphorbol Acetate; D002118:Calcium