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Calcium/calmodulin-dependent kinase kinase 2 (CaMKK2) has been implicated in a range of conditions and pathologies from prostate to hepatic cancer. Here, we describe the expression in Escherichia coli and the purification protocol for the following constructs: full-length CaMKK2 in complex with CaM, CaMKK2 'apo', CaMKK2 (165-501) in complex with CaM, and the CaMKK2 F267G mutant. The protocols described have been optimized for maximum yield and purity with minimal purification steps required and the proteins subsequently used to develop a fluorescence-based assay for drug binding to the kinase, "Using the fluorescent properties of STO-609 as a tool to assist structure-function analyses of recombinant CaMKK2" [1].

Original publication

DOI

10.1016/j.dib.2016.06.033

Type

Journal article

Journal

Data Brief

Publication Date

09/2016

Volume

8

Pages

733 - 740

Keywords

AMPK, adenosine monophosphate-regulated kinase, CaM, calmodulin, CaMK, calcium/calmodulin-dependent kinase, CaMKK2, CaMKK2, calcium/calmodulin-dependent kinase kinase 2, CaMKK2:CaM, CaMKK2-CaM complex, Calmodulin, E. coli, Escherichia coli, Fermentation, IPTG, β-D-1-thiogalactopyranoside, LB, Luria broth, LDS-PAGE, lithium dodecyl sulphate–polyacrylamide gel electrophoresis, LIC, ligation-independent cloning, OD, optical density, PMSF, phenylmethylsulfonyl fluoride, SEC, size-exclusion chromatography, TEV, tobacco etch virus, aa, amino acid