Biochemical characterization of a calcium-sensitive protein kinase LeCPK2 from tomato

文献类型： 外文期刊

第一作者： Zhang, Zhili

作者： Zhang, Zhili;Fu, Gui;Chen, Xin;Chang, Wenjun;Zhu, Jiahong

作者机构：

关键词： Calcium-dependent protein kinase;LeCPK2;Kinase-Glo (R) Luminescent Kinase Assay;Tomato

期刊名称：INDIAN JOURNAL OF BIOCHEMISTRY & BIOPHYSICS （ 影响因子：1.918； 五年影响因子：1.406 ）

ISSN： 0301-1208

年卷期： 2011 年 48 卷 3 期

页码：

收录情况： SCI

摘要： 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 Ca(2+) activation, the kinase activities of recombinant full-length and truncated LeCPK2 were measured by Kinase-Glo (R) Luminescent kinase assay (Promega). The results showed that LeCPK2 activity was Ca(2+)-dependent and the C-terminal CLD of 161 residues was essential for the activation of LeCPK2. The activity of LeCPK2 was sharply stimulated by Ca(2+) with K(0.5) (concentration of Ca(2+) for half-maximal activity) of 48.8 and 45.5 nM with substrate histone IIIs and syntide 2, respectively. The optimal concentration of Mg(2+) for LeCPK2 activity was 20 and 10 mM for substrate histone Ills and syntide 2, respectively. The K(m) value of LeCPK2 towards histone IIIs and syntide 2 was 44.9 mu g/ml and 89.52 mu M, respectively. The determination of biochemical properties of LeCPK2 would provide some clues on how its activity was regulated in vivo.

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