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In situ data collection and structure refinement from microcapillary protein crystallization

Yadav, Maneesh K. and Gerdts, Cory J. and Sanishvili, Ruslan and Smith, Ward W. and Roach, L. Spencer and Ismagilov, Rustem F. and Kuhn, Peter and Stevens, Raymond C. (2005) In situ data collection and structure refinement from microcapillary protein crystallization. Journal of Applied Crystallography, 38 (6). pp. 900-905. ISSN 0021-8898. doi:10.1107/S002188980502649X.

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In situ X-ray data collection has the potential to eliminate the challenging task of mounting and cryocooling often fragile protein crystals, reducing a major bottleneck in the structure determination process. An apparatus used to grow protein crystals in capillaries and to compare the background X-ray scattering of the components, including thin-walled glass capillaries against Teflon, and various fluorocarbon oils against each other, is described. Using thaumatin as a test case at 1.8 angstrom resolution, this study demonstrates that high-resolution electron density maps and refined models can be obtained from in situ diffraction of crystals grown in microcapillaries.

Item Type:Article
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URLURL TypeDescription CentralArticle
Ismagilov, Rustem F.0000-0002-3680-4399
Additional Information:Copyright © 2005 International Union of Crystallography. Received 1 June 2005. Accepted 18 August 2005. We are grateful to Drs Janet Smith and Robert Fischetti for the access to the GM/CA-CAT beamline at the APS during its commissioning time. GM/CA-CAT has been funded in whole or in part with Federal funds from the National Cancer Institute (Y1-CO-1020) and the National Institute of General Medical Sciences (Y1-GM-1104). Use of the APS was supported by the US Department of Energy, Office of Science, Office of Basic Energy Sciences, under Contract #W-31-109-Eng-38. This study was supported by NIH/NIAID Contract #HHSN 266200400058C, `Functional and Structural Proteomics of the SARS-CoV' (to PK), NIH Roadmap Award GM073197, `Joint Center for Innovative Membrane Protein Technology' (to RCS), NIH Protein Structure Initiative Specialized Centers Grant GM074961 (to PK, RCS and RFI), and NIH award EB001903 (to RFI). We greatly appreciate the careful reading of and comments on the manuscript by Dr Elspeth Garman.
Funding AgencyGrant Number
National Cancer InstituteY1-CO-1020
National Institute of General Medical SciencesY1-GM-1104
Department of Energy (DOE)W-31-109-ENG-38
Subject Keywords:x-ray-diffraction; microfluidic system; radiation-damage; crystallography; crystals; microbatch; resolution; diffusion; droplets; beam
Issue or Number:6
Record Number:CaltechAUTHORS:20130821-160735377
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Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:40890
Deposited By: Whitney Barlow
Deposited On:28 Aug 2013 19:32
Last Modified:10 Nov 2021 04:23

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