Remarkably Stereospecific Utilization of ATP α,β-Halomethylene Analogues by Protein Kinases

Journal of the American Chemical Society

5 years ago

Remarkably Stereospecific Utilization of ATP α,β-Halomethylene Analogues by Protein Kinases

Alvin Kung, Ming Guo, Yongheng Chen, Chao Zhang, Feng Ni, Vivek Shah, Xuegong Fan, Charles E. McKenna, Lin Chen, Yankun Duan, Carolina D. Amador

ATP analogues containing a CXY group in place of the α,β-bridging oxygen atom are powerful chemical probes for studying ATP-dependent enzymes. A limitation of such probes has been that conventional synthetic methods generate a mixture of diastereomers when the bridging carbon substitution is nonequivalent (X ≠ Y). We report here a novel method based on derivatization of a bisphosphonate precursor with a d-phenylglycine chiral auxiliary that enables preparation of the individual diastereomers of α,β-CHF-ATP and α,β-CHCl-ATP, which differ only in the configuration at the CHX carbon. When tested on a dozen divergent protein kinases, these individual diastereomers exhibit remarkable diastereospecificity (up to over 1000-fold) in utilization by the enzymes. This high selectivity can be exploited in an enzymatic approach to obtain the otherwise inaccessible diastereomers of α,β-CHBr-ATP. The crystal structure of a tyrosine kinase Src bound to α,β-CHX-ADP establishes the absolute configuration of the CHX carbon and helps clarify the origin of the remarkable diastereospecificity observed. We further synthesized the individual diastereomers of α,β-CHF-γ-thiol-ATP and demonstrated their utility in labeling a wide spectrum of kinase substrates. The novel ATP substrate analogues afforded by these two complementary strategies should have broad application in the study of the structure and function of ATP-dependent enzymes.

Publisher URL: http://dx.doi.org/10.1021/jacs.7b03266

DOI: 10.1021/jacs.7b03266