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Indian Journal of Biochemistry and Biophysics (IJBB) >
IJBB Vol.48 [2011] >
IJBB Vol.48(3) [June 2011] >
| Title: | Biochemical characterization of a calcium-sensitive protein kinase LeCPK2 from tomato |
| Authors: | Chang, Wenjun
Fu, Gui
Chen, Xin
Zhu, Jiahong
Zhang, Zhili |
| Keywords: | Calcium-dependent protein kinase
LeCPK2
Kinase-Glo® Luminescent Kinase Assay
Tomato |
| Issue Date: | Jun-2011 |
| Publisher: | NISCAIR-CSIR, India |
| 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. |
| Page(s): | 148-153 |
| CC License: |  CC Attribution-Noncommercial-No Derivative Works 2.5 India |
| ISSN: | 0975-0959 (Online); 0301-1208 (Print) |
| Source: | IJBB Vol.48(3) [June 2011]
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