Development of the procedures for high-yield expression and rapid purification of active recombinant Csk-homologous kinase (CHK): Comparison of the catalytic activities of CHK and CSK
Khai-Chew Chan, Daisy Sio-Seng Lio, Renwick CJ Dobson, Boonyarin Jarasrassamee, Mohammed I Hossain, Aainaa K Roslee, Kim K Ia, Matthew A Perugini, Heung-Chin Cheng
PROTEIN EXPRESSION AND PURIFICATION | ACADEMIC PRESS INC ELSEVIER SCIENCE | Published : 2010
Csk-homologous kinase (CHK) is an important endogenous inhibitor constraining the oncogenic actions of Src-family kinases (SFKs) in cells. It suppresses SFK activity by specifically phosphorylating the conserved regulatory tyrosine near the C-terminus of SFKs. In addition to phosphorylation, CHK employs a novel non-catalytic inhibitory mechanism to suppress SFK activity. This mechanism involves direct binding of CHK to the active forms of SFKs to form stable protein complexes. Since aberrant activation of SFKs contributes to cancer formation and progression, small-molecule inhibitors mimicking the non-catalytic inhibitory mechanism of CHK are potential anti-cancer therapeutics. Elucidation o..View full abstract
The works described in this manuscript were supported by project grants of the National Health and Medical Research Council of Australia and the Cancer Council Victoria. We wish to thank Dr. David Morgan of University of California, San Francisco for his generous gift of the recombinant CSK-baculovirus. We wish to thank Dr. Nick Williamson of the mass spectrometry and peptide synthesis core facility of the Bio21 Institute for help in molecular weight determination of the recombinant proteins by mass spectrometry. M.P.P acknowledges the Australian Research Council (ARC) for Future Fellowship support. R.C.J.D. is supported by the CR Roper Fellowship of the Faculty of Medicine, Dentistry and Health Sciences, University of Melbourne.