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Successful Flash-Cooling of Xenon Derivatized Myoglobin Crystals

Soltis, S. Michael and Stowell, Michael H. B. and Wiener, Michael C. and Phillips, George N., Jr. and Rees, D. C. (1997) Successful Flash-Cooling of Xenon Derivatized Myoglobin Crystals. Journal of Applied Crystallography, 30 (2). pp. 190-194. ISSN 0021-8898. doi:10.1107/S0021889896012939.

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This paper demonstrates for the first time a method for preparing cryocooled xenon-derivatized protein crystals. The method is based upon the hypothesis and subsequent observation that the diffusion of a xenon atom from a tight binding site following depressurization occurs on a timescale of minutes. We have observed significant changes in diffraction intensities from myoglobin crystals for up to 5 min following depressurization from 1 MPa of xenon. In accordance with this observation, a xenon-derivatized myoglobin crystal was cryocooled at ~95 K within 20 s of complete depressurization. A crystallographic data set was then collected to 2.0 Å resolution and isomorphous and anomalous difference Patterson maps revealed the presence of a well ordered xenon site with an occupancy of approximately 0.5. Phasing statistics for this site were of good quality and demonstrate the practicality of this method. The ability to cryocool xenon-derivatized crystals will make this heavy-atom substitution method even more useful for single-isomorphous-replacement and multiple-isomorphous-replacement phasing of macromolecules.

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Rees, D. C.0000-0003-4073-1185
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Additional Information:© 1997 International Union of Crystallography. Received 18 July 1996, accepted 16 October 1996. This work was supported in part by NIH GM45062 of the NSF to DCR and by the DOE, Office of Basic Energy Sciences and the Office of Health and Environmental Research, and by the NIH, Biomedical Research Technology Program, National Center for Research Resources for support of the rotation camera facilities at SSRL. We thank R. P. Phizackerley and Henry D. Bellamy for helpful discussions.
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Department of Energy (DOE)UNSPECIFIED
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ID Code:54433
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Deposited On:08 Feb 2015 03:28
Last Modified:10 Nov 2021 20:33

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