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Int. J. Mol. Sci. 2013, 14(12), 24501-24513; doi:10.3390/ijms141224501
Article

Identification and Biochemical Characterization of Protein Phosphatase 5 from the Cantharidin-Producing Blister Beetle, Epicauta chinensis

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Received: 28 November 2013; in revised form: 9 December 2013 / Accepted: 11 December 2013 / Published: 16 December 2013
(This article belongs to the Section Biochemistry, Molecular Biology and Biophysics)
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Abstract: Protein phosphatase 5 (PP5) is a unique member of serine/threonine phosphatases which has been recognized in regulation of diverse cellular processes. A cDNA fragment encoding PP5 (EcPP5) was cloned and characterized from the cantharidin-producing blister beetle, E. chinensis. EcPP5 contains an open reading frame of 1500 bp that encodes a protein of 56.89 kDa. The deduced amino acid sequence shares 88% and 68% identities to the PP5 of Tribolium castaneum and humans, respectively. Analysis of the primary sequence shows that EcPP5 has three TPR (tetratricopeptide repeat) motifs at its N-terminal region and contains a highly conserved C-terminal catalytic domain. RT-PCR reveals that EcPP5 is expressed in all developmental stages and in different tissues. The recombinant EcPP5 (rEcPP5) was produced in Escherichia coli and purified to homogeneity. The purified protein exhibited phosphatase activity towards pNPP (p-nitrophenyl phosphate) and phosphopeptides, and its activity can be enhanced by arachidonic acid. In vitro inhibition study revealed that protein phosphatase inhibitors, okadaic acid, cantharidin, norcantharidin and endothall, inhibited its activity. Further, protein phosphatase activity of total soluble protein extract from E. chinensis adults could be impeded by these inhibitors suggesting there might be some mechanism to protect this beetle from being damaged by its self-produced cantharidin.
Keywords: Epicauta chinensis; protein phosphatase 5; E. coli expression; in vitro inhibition Epicauta chinensis; protein phosphatase 5; E. coli expression; in vitro inhibition
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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MDPI and ACS Style

Chen, X.; Lü, S.; Zhang, Y. Identification and Biochemical Characterization of Protein Phosphatase 5 from the Cantharidin-Producing Blister Beetle, Epicauta chinensis. Int. J. Mol. Sci. 2013, 14, 24501-24513.

AMA Style

Chen X, Lü S, Zhang Y. Identification and Biochemical Characterization of Protein Phosphatase 5 from the Cantharidin-Producing Blister Beetle, Epicauta chinensis. International Journal of Molecular Sciences. 2013; 14(12):24501-24513.

Chicago/Turabian Style

Chen, Xi'en; Lü, Shumin; Zhang, Yalin. 2013. "Identification and Biochemical Characterization of Protein Phosphatase 5 from the Cantharidin-Producing Blister Beetle, Epicauta chinensis." Int. J. Mol. Sci. 14, no. 12: 24501-24513.


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