Trypanosoma cruzi epimastigote forms possess a Ca 2+-calmodulin dependent protein kinase

• Published in: FEBS letters Vol. 337; no. 3; pp. 293 - 297
• Main Authors: Ogueta, S.B., Solari, A., Téllez-Iñón, M.T.
• Format: Journal Article
• Language: English
• Published: Elsevier B.V 17.01.1994
• Subjects: Ca 2+-calmodulin protein kinase; Cytoskeleton; Flagella; Trypanosoma cruzi; Trypanosomatid; ATP, adenosine 5'-triphosphate; SDS-PAGE, sodium dodecyl sulfate-polyacrylamide gel electrophoresis; TLCK, Nα- p-tosyl- l-lysine chloro-methyl ketone; CaM, calmodulin; Trypanosoma cruzi; cAMP, cyclic adenosine monophosphate; CaM kinase II, Ca 2+-calmodulin-dependent protein kinase type II; Trypanosomatid; DTT, d, l-dithiothreitol; EDTA, ethylendiaminetetraacetic acid; βME, β-mercaptoethanol; E 64 trans-epoxysuccinyl- l-leucylamido (4-guanidino) butane; PMSF, phenylmethyl-sulfonyl fluoride; Ca 2+-calmodulin protein kinase; Cytoskeleton; Flagella; EGTA, ethylene glycol-bis(β-amino-ethylether)
• ISSN: 0014-5793; 1873-3468
• DOI: 10.1016/0014-5793(94)80212-2

Trypanosoma cruzi epimastigote forms showed a tightly bound Ca 2+-calmodulin-dependent protein kinase activity, which could be partially extracted from membranes and axonemes. The enzyme is constituted by subunits which were autophosphorylated in the absence of exogenous substrates. An antibody against CaM kinase II recognized a Ca 2+- or Ca 2+-CaM-dependent conformational epitope in these fractions. The detected bands were of molecular weights similar to the α and β subunits of the corresponding bovine brain enzyme (60 and 50 kDa). Studies using [ 125I]CaM revealed the presence of a CaM-binding domain. These experiments confirm that the parasite possesses a paniculate CaM kinase with characteristics similar to the bovine brain enzyme.