Abstract
Total synthesis of proteins by modern chemical ligation methods enables the ready preparation of high purity protein molecules of typical size (up to ∼300 amino acid residues). This in turn enables the preparation of mirror image D-protein molecules not found in nature. Use of a racemic protein mixture (i.e. D-protein+L-protein) greatly facilitates the formation of diffraction-quality crystals of otherwise recalcitrant proteins. Facilitated crystallization is also observed for quasi-racemic protein mixtures. Centrosymmetric crystals of racemic proteins diffract to high resolution and offer enhanced possibilities for structure solution by direct computational methods. Racemic protein crystallography has been successfully applied to a number of recalcitrant protein molecules, and has been used to determine the structure of a 35kDa {L-protein target/D-protein ligand} complex.
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Acknowledgments
This research was supported by the Office of Science (BER), U.S. Department of Energy (grant no. DE-FG02 07ER64501 to S.B.H.K.) and by the National Institutes of Health (grant no. R01 GM075993 to S.B.H.K.). Use of NE-CAT beamline 24-ID at the Advanced Photon Source is supported by award RR-15301 from the National Center for Research Resources at the National Institutes of Health. Use of the Advanced Photon Source is supported by the U.S. Department of Energy, Office of Basic Energy Sciences, under contract no. DE-AC02-06CH11357.
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Mandal, K., Kent, S.B.H. (2013). Total Chemical Protein Synthesis for the Determination of Novel X-ray Structures by Racemic Protein Crystallography. In: Read, R., Urzhumtsev, A., Lunin, V. (eds) Advancing Methods for Biomolecular Crystallography. NATO Science for Peace and Security Series A: Chemistry and Biology. Springer, Dordrecht. https://doi.org/10.1007/978-94-007-6232-9_2
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