Data for the co-expression and purification of human recombinant CaMKK2 in complex with calmodulin in Escherichia coli

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,...

Descripción completa

Detalles Bibliográficos
Autores principales: Gerner, Lisa, Munack, Steffi, Temmerman, Koen, Lawrence-Dörner, Ann-Marie, Besir, Hüseyin, Wilmanns, Matthias, Jensen, Jan Kristian, Thiede, Bernd, Mills, Ian G., Morth, Jens Preben
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2016
Materias:
Data Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4950174/
https://www.ncbi.nlm.nih.gov/pubmed/27508226
http://dx.doi.org/10.1016/j.dib.2016.06.033
Descripción
Sumario: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].

Ejemplares similares