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An optimal exposure strategy for cryoprotected virus crystals with lattice constants greater than 1000 Å

Gan, L. and Johnson, J. E. (2008) An optimal exposure strategy for cryoprotected virus crystals with lattice constants greater than 1000 Å. Journal of Synchrotron Radiation, 15 (3). pp. 223-226. ISSN 0909-0495. doi:10.1107/S0909049507064813.

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Studies of icosahedral virus capsids provide insights into the function of supramolecular machines. Virus capsid crystals have exceptionally large unit cells; as a result, they diffract weakly compared with protein crystals. HK97 is a dsDNA lambda-like bacteriophage whose 13 MDa capsid expands from 550 Å to 650 Å with large subunit conformational changes during virus maturation. The HK97 penultimate maturation intermediate was crystallized in a tetragonal unit cell that has lattice constants of 1010 Å × 1010 Å × 730 Å. The crystals could be cryoprotected, but diffracted to a modest resolution of 5 Å at a bending-magnet beamline. When these crystals were optimally exposed with two orders-of-magnitude more photons from a new insertion-device beamline, data extending to better than 3.8 Å resolution were obtained. Here, the strategies to collect and process such data are described. These strategies can be adapted for other crystals with large unit cells and for microcrystals.

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Additional Information:© 2008 International Union of Crystallography. Received 3 August 2007. Accepted 30 November 2007. We thank our many colleagues, especially James Conway, Robert Duda, Ilya Gertsman, Roger Hendrix, Gabriel Lander, Kelly Lee, Lars Liljas, Tianwei Lin, Vijay Reddy, Jeffrey Speir and Alasdair Steven for stimulating discussions. We thank the staff of beamlines 11-1 of SSRL, and 14-BMC and 23-ID-D of APS for assistance in data collection. In particular, we thank Spencer Anderson, Keith Brister, Robert Fischetti, Nukri Sanishvili and Ward Smith for advising and helping with data collection. Research carried out at the SSRL was supported by the DOE, OBER and by the NIH, NCRR, Biomedical Technology Program, and the NIGMS. Research carried out at the APS was supported by the US DOE Contract No. W-31-109-Eng-38, research carried out at GM/CA CAT was supported by the NCI (Y1- CO-1020) and the NIGMS (Y1-GM-1104), and research carried out at BioCARS sector 14 was supported by the NIH, NCRR, under grant RR07707. LG was supported by a Fletcher Jones Foundation Fellowship. This work was supported by NIH grant R01 AI40101 to JEJ.
Funding AgencyGrant Number
Department of EnergyW-31-109-Eng-38
National Cancer InstituteY1-CO-1020
National Institute of General Medical SciencesY1-GM-1104
National Institute of HealthRR07707
National Center for Research ResourcesRR07707
National Institute of HealthR01 AI40101
Fletcher Jones FoundationUNSPECIFIED
Subject Keywords:virus crystals; bacteriophage HK97; insertion-device beamlines
Issue or Number:3
Record Number:CaltechAUTHORS:GANjsr08
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Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:13661
Deposited By: Tony Diaz
Deposited On:19 Jun 2009 18:27
Last Modified:08 Nov 2021 22:39

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