title: Data for the co-expression and purification of human recombinant CaMKK2 in complex with calmodulin in Escherichia coli.
abstract: 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].
authors: Centre for Molecular Medicine Norway (NCMM), Nordic EMBL Partnership, Forskningsparken, University of Oslo and Oslo University Hospitals, 0349 Oslo, Norway.
authors: Lisa
authors: L
authors: Gerner
authors: Centre for Molecular Medicine Norway (NCMM), Nordic EMBL Partnership, Forskningsparken, University of Oslo and Oslo University Hospitals, 0349 Oslo, Norway.
authors: Steffi
authors: S
authors: Munack
authors: European Molecular Biology Laboratory Hamburg, Notkestrasse 85, 22603 Hamburg, Germany; European Molecular Biology Laboratory Heidelberg, Meyerhofstrasse 1, 69117 Heidelberg, Germany.
authors: Koen
authors: K
authors: Temmerman
authors: European Molecular Biology Laboratory Heidelberg, Meyerhofstrasse 1, 69117 Heidelberg, Germany; Institute of Biochemistry, WWU Münster, Wilhelm-Klemm-Straße 2, 48149 Münster, Germany.
authors: Ann-Marie
authors: AM
authors: Lawrence-Dörner
authors: European Molecular Biology Laboratory Heidelberg, Meyerhofstrasse 1, 69117 Heidelberg, Germany.
authors: Hüseyin
authors: H
authors: Besir
authors: European Molecular Biology Laboratory Hamburg, Notkestrasse 85, 22603 Hamburg, Germany.
authors: Matthias
authors: M
authors: Wilmanns
authors: Department of Molecular Biology and Genetics, Aarhus University, 8000 Aarhus, Denmark.
authors: Jan Kristian
authors: JK
authors: Jensen
authors: Department of Biosciences, University of Oslo, Norway; Biotechnology Centre of Oslo, University of Oslo, Norway.
authors: Bernd
authors: B
authors: Thiede
authors: Centre for Molecular Medicine Norway (NCMM), Nordic EMBL Partnership, Forskningsparken, University of Oslo and Oslo University Hospitals, 0349 Oslo, Norway; Department of Molecular Oncology, Institute for Cancer Research, Oslo University Hospitals, Norway; Movember/Prostate Cancer UK Centre of Excellence for Prostate Cancer Research, Centre for Cancer Research and Cell Biology (CCRCB), Queen׳s University Belfast, UK.
authors: Ian G
authors: IG
authors: Mills
authors: Centre for Molecular Medicine Norway (NCMM), Nordic EMBL Partnership, Forskningsparken, University of Oslo and Oslo University Hospitals, 0349 Oslo, Norway; Institute for Experimental Medical Research, Oslo University Hospital, Norway.
authors: Jens Preben
authors: JP
authors: Morth
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