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X-ray structure determination of the glycine cleavage system protein H of Mycobacterium tuberculosis using an inverse Compton synchrotron X-ray source

  • Published:
Journal of Structural and Functional Genomics

Abstract

Structural genomics discovery projects require ready access to both X-ray diffraction and NMR spectroscopy which support the collection of experimental data needed to solve large numbers of novel protein structures. The most productive X-ray crystal structure determination laboratories make extensive use of tunable synchrotron X-ray light to solve novel structures by anomalous diffraction methods. This requires that frozen cryo-protected crystals be shipped to large multi acre synchrotron facilities for data collection. In this paper we report on the development and use of the first laboratory-scale synchrotron light source capable of performing many of the state-of-the-art synchrotron applications in X-ray science. This Compact Light Source is a first-in-class device that uses inverse Compton scattering to generate X-rays of sufficient flux, tunable wavelength and beam size to allow high-resolution X-ray diffraction data collection from protein crystals. We report on benchmarking tests of X-ray diffraction data collection with hen egg white lysozyme, and the successful high-resolution X-ray structure determination of the Glycine cleavage system protein H from Mycobacterium tuberculosis using diffraction data collected with the Compact Light Source X-ray beam.

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Acknowledgments

We gratefully acknowledge NIH funding for the developments included in this paper. The prototype CLS was funded by the NIGMS under grant R44-GM66511. The X-ray station and optics development was funded by the NIGMS and NCRR under grant R44-GM074437. The development of the Beta CLS was funded by the NIGMS and NCRR which have co-sponsored the Accelerated Technologies Center for Gene to 3D Structure (www.atcg3d.org), a PSI-2 Specialized Center, under Grant U54 GM074961. The production of MytuGCSPH protein and crystals was funded by NIAID under Federal Contract No. HHSN272200700057C which supports the Seattle Structural Genomics Center for Infectious Disease (www.SSGCID.org). Special thanks to Shellie Dieterich, Becky Poplawski, and Jeff Christensen at Emerald BioStructures for their support in molecular biology and protein crystallization. The authors also wish to thank the organizing committee of the PSI organized Enabling Technologies Meetings held each spring on the NIH campus. LTI collaborators wish to thank the staff at LTI, Michael Blum, Jens Als-Nielsen, Anette Jensen and Bill Weis.

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Authors and Affiliations

  1. Emerald BioStructures, 7869 NE Day Road West, Bainbridge Island, WA, 98110, USA

    Jan Abendroth, Thomas E. Edwards, Bart Staker & Lance J. Stewart

  2. The Scripps Research Institute, 10550 N. Torrey Pines Road, La Jolla, CA, 92037, USA

    Michael S. McCormick & Peter Kuhn

  3. Lyncean Technologies, 370 Portage Ave, Palo Alto, CA, 94306, USA

    Roderick Loewen, Martin Gifford, Jeff Rifkin & Ronald D. Ruth

  4. Seattle Biomedical Research Institute, 307 Westlake Ave N., Suite 500, Seattle, WA, 98109, USA

    Chad Mayer, Wenjin Guo, Yang Zhang & Peter Myler

  5. University of Washington, Seattle, WA, 98195, USA

    Angela Kelley, Erwin Analau, Stephen Nakazawa Hewitt & Alberto J. Napuli

  6. SLAC National Accelerator Laboratory, Stanford, CA, 94305, USA

    Ronald D. Ruth

Authors
  1. Jan Abendroth
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  2. Michael S. McCormick
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  3. Thomas E. Edwards
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  4. Bart Staker
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  5. Roderick Loewen
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  6. Martin Gifford
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  7. Jeff Rifkin
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  8. Chad Mayer
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  9. Wenjin Guo
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  10. Yang Zhang
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  11. Peter Myler
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  12. Angela Kelley
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  13. Erwin Analau
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  14. Stephen Nakazawa Hewitt
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  15. Alberto J. Napuli
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  16. Peter Kuhn
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  17. Ronald D. Ruth
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  18. Lance J. Stewart
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Corresponding authors

Correspondence to Ronald D. Ruth or Lance J. Stewart.

Additional information

Lance J. Stewart is corresponding author for the protein structure research aspects of the publication and Ronald D. Ruth is corresponding author for the design and function of the CLS aspects of the publication.

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Abendroth, J., McCormick, M.S., Edwards, T.E. et al. X-ray structure determination of the glycine cleavage system protein H of Mycobacterium tuberculosis using an inverse Compton synchrotron X-ray source. J Struct Funct Genomics 11, 91–100 (2010). https://doi.org/10.1007/s10969-010-9087-6

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  • DOI: https://doi.org/10.1007/s10969-010-9087-6

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