Biochemical characterization of a calcium-sensitive protein kinase LeCPK2 from tomato.
Indian J Biochem Biophys
;
2011 June; 48(3): 148-153
Artículo
en Inglés
| IMSEAR
| ID: sea-135312
ABSTRACT
LeCPK2 (GenBank GQ205414), a versatile calcium-dependent protein kinase (CDPK or CPK) gene was isolated from tomato in our previous study. In this study, the biochemical properties of LeCPK2 were further investigated. To examine the role of the C-terminal calmodulin-like domain (CLD) of LeCPK2 with respect to Ca2+ activation, the kinase activities of recombinant full-length and truncated LeCPK2 were measured by Kinase-Glo® Luminescent kinase assay (Promega). The results showed that LeCPK2 activity was Ca2+-dependent and the C-terminal CLD of 161 residues was essential for the activation of LeCPK2. The activity of LeCPK2 was sharply stimulated by Ca2+ with K0.5 (concentration of Ca2+ for half-maximal activity) of 48.8 and 45.5 nM with substrate histone IIIs and syntide 2, respectively. The optimal concentration of Mg2+ for LeCPK2 activity was 20 and 10 mM for substrate histone IIIs and syntide 2, respectively. The Km value of LeCPK2 towards histone IIIs and syntide 2 was 44.9 μg/ml and 89.52 μM, respectively. The determination of biochemical properties of LeCPK2 would provide some clues on how its activity was regulated in vivo.
Texto completo:
Disponible
Índice:
IMSEAR (Asia Sudoriental)
Asunto principal:
Proteínas Quinasas
/
Especificidad por Sustrato
/
Proteínas Recombinantes
/
Cloruro de Calcio
/
Datos de Secuencia Molecular
/
Cloruro de Magnesio
/
Secuencia de Aminoácidos
/
Solanum lycopersicum
Idioma:
Inglés
Revista:
Indian J Biochem Biophys
Año:
2011
Tipo del documento:
Artículo
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