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].

CaMKK2 facilitates Golgi-associated vesicle trafficking to sustain cancer cell proliferation

“…pGEX6P-1 constructs for CaMKK2 isoform 2 and Gemin4 (aa 1–370) were expressed and purified as previously described [ 20 ].…”

A Nuclease from Streptococcus mutans Facilitates Biofilm Dispersal and Escape from Killing by Neutrophil Extracellular Traps

“…A single colony was inoculated into LB medium and cultured at 16°C after the addition of 0.3 mM (final concentration) isopropyl-β-D-thiogalactopyranoside to induce protein expression. The overnight culture was collected for recombinant DeoC protein purification as described previously (Gerner et al, 2016). In brief, the cell pellets were resuspended in lysis buffer (20 mM Tris, 500 mMNaCl, pH 8.0), lysed using an Ultrasonic Cell Disruptor (VCX130, 3 mm; Sonics, USA) for 120 × 4 s and cellular debris was removed by centrifugation at 12,000 g for 25 min at 4°C.…”