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Study of protein kinases CDK8 and CDK10 from heterologous expression in E. coli

Grant number: 16/25893-6
Support type:Regular Research Grants
Duration: April 01, 2017 - March 31, 2019
Field of knowledge:Biological Sciences - Biochemistry - Chemistry of Macromolecules
Principal researcher:Fernanda Canduri
Grantee:Fernanda Canduri
Home Institution: Instituto de Química de São Carlos (IQSC). Universidade de São Paulo (USP). São Carlos , SP, Brazil


Protein kinases are important targets for study due to their involvement in cell signaling, transcription and cell division. Cyclin dependent kinases (CDKs) comprise a family of proteins that can be subdivided into two major groups based on their functional role in cell cycle and/or transcriptional control. The complexes of the CDK10 associate with the ETS2 transcription factor, and they are involved with RNA splicing, regulation of steroid hormones, as well as participate in other transcriptional regulatory complexes. The CDK8 has a motif mediators binding, and therefore is part of the complex of transcriptional mediator. In this proposal, the CDK8 and CDK10 will be overexpressed and purified following the protocol established already, and functional and structural studies will be carried out in order to achieve greater understanding of the differential with respect to structure-function of each. The structural studies will be performed using circular dichroism methods and methods of protein crystallization in an attempt to obtain monocrystals to be used in X-ray diffraction and resolution structure. In addition, molecular docking studies will be performed in the search for ATP-competitive ligand that will guide in vitro studies of these CDKs in the presence of different ligands previously selected, using UV/Vis spectrophotometric methods. These studies may help to understand the mode of interaction of CDK8 and CDK10 with ATP competitive ligands, as well as to develop chemical inhibitors of low molecular weight which can specifically inhibit these kinases. (AU)

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Scientific publications
(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)
COELHO, FERNANDA; PEPINO, REBEKA DE OLIVEIRA; ALENCAR, DIANDRA PINHEIRO; SANTOS, JESSYKA LIMA; CANDURI, FERNANDA. Overexpression and refolding of human Cyclin D3. A reliable method or not?. Process Biochemistry, v. 99, p. 196-201, DEC 2020. Web of Science Citations: 0.

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