Abl kinase constructs expressed in bacteria: facilitation of structural and functional studies including segmental labeling by expressed protein ligation

摘要: A great portion of tyrosine kinases are involved in cell development and their structural alteration is intimately associated pathologies oncology. These one the major groups targets under investigation for molecular therapeutics. To carry out biochemical biological studies on these kinases, economical production purified forms highly desirable. However over-expressing as recombinant bacterial systems purification a significant challenge. Abelson kinase (Abl) has previously been expressed large scale to facilitate X-ray crystallography NMR structure mainly baculovirus infected insect cells. Even though success achieved expression soluble E. coli with chaperones improve correct folding, low levels intrinsic such because diversion resources produce chaperones. Here we present straightforward method express purify isolated Abl domain SH3–SH2–kinase multi-domain structures. The protein retains its folding function. yield several mg L−1 range minimal media. Furthermore demonstrate that segmental isotopic labelling using ligation can be constructs, which especially useful structure–activity studies.