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4.7. Selection of optimal ligand-induced protein conformations
Each ligand-induced protein conformation was analysed by
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4.8. Protein production and crystallography
The CHK2 kinase domain (CHK2-KD, amino acids 210-531) was
produced as a GST-fusion protein and purified as previously de-
scribed.42 Co-crystallisation experiments with four selected benz-
imidazole inhibitors were carried out based upon conditions
described earlier.12,13,42 For full details of these experiments, crys-
tallographic data collection and refinement see Supplementary
data and Table S6.
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4.9. Chemistry methods
Compound structures have previously been disclosed although
preparative methods have not been described for all com-
pounds.15–17 Materials and methods used to prepare compounds
19, 30, 44 and 47 for protein–ligand crystallography are described
in Supplementary data.
4.10. Accession codes
Atomic coordinates and structure factors for the crystal struc-
tures of ligand-bound CHK2 can be accessed using the following
PDB codes: 19, 4A9S; 30, 4A9R; 44, 4A9T and 47, 4A9U.
Acknowledgments
We acknowledge NHS funding to the NIHR Biomedical Research
Centre. I.M.W. and R.v.M. are supported by The Institute of Cancer
Research and Cancer Research UK Grant Number C309/A8274; J.B.
is supported by Cancer Research UK Grant Number C309/A8274.
M.C.S.-S. is supported by a Wellcome Trust doctoral training pro-
gram. We thank Dr. Nathan Brown, Dr. Michelle Garrett and Dr.
Ian Collins for their valuable comments and helpful discussions.
We are grateful to Dr. Maggie Liu for collection and interpretation
of the compound NMR spectra. We thank Dr. Antony Oliver and
Prof Laurence Pearl (University of Sussex, formerly ICR) for kindly
donating the CHK2 crystallography construct and for helpful dis-
cussions. We also thank the staff of the DIAMOND light source
beamline I03 for their support during data collection.
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Supplementary data
Supplementary data (the ligands used in the flexible docking
study Table S1), the residue selection for side chain flexible dock-
ing (Tables S2–S4) and experimental protocols for preparation of
compounds studied by co-crystallography and associated crystal-
lographic analysis associated with this article can be found in the
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References and notes
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