Following ligand binding and ligand reactions in proteins via Raman crystallography

Biochemistry
Paul R Carey, Jian Dong

Abstract

Raman crystallography permits the monitoring of chemical events in single-protein crystals in real time. Using a Raman microscope, it is possible to obtain protein Raman spectroscopic data of unprecedented quality and stability. The latter features allow us to obtain the Raman spectrum for small molecules soaking into crystals under normal (nonresonance) Raman conditions. Thus, via an approach utilizing Raman difference spectroscopy, we can quantitate the amount of ligand in the crystal, determine the chemistry of inhibitor-protein interactions, and follow chemical reactions in the active site on the time scale of minutes. While providing unique chemical insights, these data also provide an invaluable guide for determining the conditions for flash-freezing crystals for X-ray crystallographic analysis. In addition, the Raman difference spectra often contain contributions from protein modes due to protein conformational changes occurring upon ligand binding. These features allow us to probe events ranging from small cooperative conformational changes to massive and unexpected secondary structure changes in the crystal. An experimental advantage of Raman crystallography is that the data can be collected from crystals in situ, in s...Continue Reading

References

Jan 1, 1993·Methods in Enzymology·G Smulevich, T G Spiro
Sep 10, 1999·The Journal of Biological Chemistry·P R Carey

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Citations

Nov 6, 2010·Nucleic Acids Research·Jin Ok YangSanghyuk Lee
Feb 26, 2009·Journal of Synchrotron Radiation·Robin L OwenClemens Schulze-Briese
Apr 8, 2006·Annual Review of Physical Chemistry·Paul R Carey
Sep 12, 2007·Journal of Pharmaceutical Sciences·Zai-Qing Wen
Aug 10, 2010·Biochimica Et Biophysica Acta·John E McGeehanPhilippe Carpentier
Aug 28, 2010·Biochimica Et Biophysica Acta·Paul R CareyMatthew Kalp
May 5, 2009·Methods : a Companion to Methods in Enzymology·Bo GongPhilip C Bevilacqua
Jul 8, 2008·Biophysical Chemistry·Antonello MerlinoAlessandro Vergara
Aug 15, 2015·Annals of the New York Academy of Sciences·Paul R Carey, Hossein Heidari-Torkabadi
Nov 24, 2005·Journal of Molecular Biology·Mihaela M ApetriVernon E Anderson
Sep 1, 2005·Current Opinion in Structural Biology·Dominique Bourgeois, Antoine Royant
Nov 21, 2007·Analytical Biochemistry·Yong XieDor Ben-Amotz
Feb 22, 2007·Applied Spectroscopy·Kohki NodaHideo Tashiro
Aug 10, 2006·Chemical Reviews·Paul R Carey

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