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Contemporary Use of Anomalous Diffraction in Biomolecular Structure Analysis

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Protein Crystallography

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1607))

Abstract

The normal elastic X-ray scattering that depends only on electron density can be modulated by an “anomalous” component due to resonance between X-rays and electronic orbitals. Anomalous scattering thereby precisely identifies atomic species, since orbitals distinguish atomic elements, which enables the multi- and single-wavelength anomalous diffraction (MAD and SAD) methods. SAD now predominates in de novo structure determination of biological macromolecules, and we focus here on the prevailing SAD method. We describe the anomalous phasing theory and the periodic table of phasing elements that are available for SAD experiments, differentiating between those readily accessible for at-resonance experiments and those that can be effective away from an edge. We describe procedures for present-day SAD phasing experiments and we discuss optimization of anomalous signals for challenging applications. We also describe methods for using anomalous signals as molecular markers for tracing and element identification. Emerging developments and perspectives are discussed in brief.

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Acknowledgments

This work was supported in part by NIH grants R01GM107462 and P41GM116799 and by Brookhaven National Laboratory LDRD 15-034.

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Authors and Affiliations

  1. Biology Department, Brookhaven National Laboratory, PO Box 5000, 50 Bell Ave, Building 463, Upton, NY, 11973, USA

    Qun Liu

  2. Department of Biochemistry and Molecular Biophysics, Columbia University, 202 Black Building, 650 West 168th Street, New York, NY, 10032, USA

    Wayne A. Hendrickson

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  1. Qun Liu
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  2. Wayne A. Hendrickson
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Correspondence to Qun Liu or Wayne A. Hendrickson .

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Editors and Affiliations

  1. Macromolecular Crystallography Laboratory, National Cancer Institute, Frederick, Maryland, USA

    Alexander Wlodawer

  2. Synchrotron Radiation Research Section, MCL, National Cancer Institute, Argonne National Laboratory, Argonne, Illinois, USA

    Zbigniew Dauter

  3. Department of Crystallography, Faculty of Chemistry, A. Mickiewicz University, Poznan, Poland

    Mariusz Jaskolski

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Liu, Q., Hendrickson, W.A. (2017). Contemporary Use of Anomalous Diffraction in Biomolecular Structure Analysis. In: Wlodawer, A., Dauter, Z., Jaskolski, M. (eds) Protein Crystallography. Methods in Molecular Biology, vol 1607. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7000-1_16

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  • DOI: https://doi.org/10.1007/978-1-4939-7000-1_16

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