Efficient soluble expression of active recombinant human cyclin A2 mediated by E. coli molecular chaperones.

ABSTRACT: Bacterial expression of human proteins continues to present a critical challenge in protein crystallography and drug design. While human cyclin A constructs have been extensively characterized in complex with cyclin dependent kinase 2 (CDK2), efforts to express the monomeric human cyclin A2 in Escherichia coli in a stable form, without the kinase subunit, have been laden with technical difficulties, including solubility, yield and purity. Here, optimized conditions are described with the aim of generating for first time, sufficient quantities of human recombinant cyclin A2 in a soluble and active form for crystallization and ligand characterization purposes. The studies involve implementation of a His-tagged heterologous expression system under conditions of auto-induction and mediated by

SUBMITTER: Grigoroudis AI 

PROVIDER: S-EPMC4456239 | biostudies-literature | 2015 Sep

REPOSITORIES: biostudies-literature