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

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• 作者：Lisa Gerner^a ; Steffi Munack^a ; Koen Temmerman^b ; ^c ; Ann-Marie Lawrence-Dö ; rner^c ; ^d ; Hü ; seyin Besir^c ; Matthias Wilmanns^b ; Jan Kristian Jensen^e ; Bernd Thiede^f ; ^g ; Ian G. Mills^a ; ^h ; ⁱ ; ¹ ; ^{ian.mills@ncmm.uio.no" class="auth_mail" title="E-mail the corresponding author} ; ^{i.mills@qub.ac.uk" class="auth_mail" title="E-mail the corresponding author} ; Jens Preben Morth^a ; ^j ; ¹ ; ^{j.p.morth@ncmm.uio.no" class="auth_mail" title="E-mail the corresponding author}
• 关键词：aa ; amino acid ; AMPK ; adenosine monophosphate-regulated kinase ; CaM ; calmodulin ; CaMK ; calcium/calmodulin-dependent kinase ; CaMKK2 ; calcium/calmodulin-dependent kinase kinase 2 ; CaMKK2 ; CaM ; CaMKK2-CaM complex ; E. coli ; Escherichia coli ; IPTG ; β-D-1-thiogalactopyranoside ; LB ; Luria broth ; LDS-PAGE ; lithium dodecyl sulphate&ndash ; polyacrylamide gel electrophoresis ; LIC ; ligation-independent cloning ; OD ; optical density ; PMSF ; phenylmethylsulfonyl fluoride ; SEC ; size-exclusion chromatography ; TEV
• 刊名：Data in Brief
• 出版年：2016
• 出版时间：September 2016
• 年：2016
• 卷：8
• 期：Complete
• 页码：733-740
• 全文大小：1248 K

文摘

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].