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High-Throughput Studies of Protein Shapes and Interactions by Synchrotron Small-Angle X-Ray Scattering

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Structural Proteomics

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1261))

Abstract

Solution-based small angle X-ray scattering (SAXS) affords the opportunity to extract accurate structural parameters and global shape information from diverse biological macromolecular systems. SAXS is an ideal complementary technique to other structural and biophysical methods but it can also be applied alone to access structural information that is otherwise unobtainable using high-resolution methods. Macromolecular structures ranging from kilodaltons to gigadaltons can be analyzed, which encompasses the size of most proteins and functional cellular complexes. The SAXS analysis is performed using only a few microliters of solution containing microgram quantities of purified material in sample environments that can be tailored to mimic physiological conditions or altered to suit a particular question. High-brilliance synchrotron X-ray sources and parallel advances in hardware and computing have reduced data acquisition times to the millisecond range and the application of automated methods have allowed data processing and low resolution shape modelling to be completed within minutes. These developments have paved the way for high-throughput studies that generate significant quantities of structural information over a short period of time. Here, we briefly consider the basics of SAXS and describe major methods and protocols employed in high-throughput SAXS studies.

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Acknowledgements

We thank Dr David Jacques of the MRC Laboratory of Molecular Biology, Cambridge, for measuring the SAXS data presented in Figures 4 and 5. This work was supported by the Bundesministerium für Bildung und Forschung (BMBF) project BIOSCAT [05K12YE1].

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Authors and Affiliations

  1. European Molecular Biology Laboratory (EMBL), Hamburg Outstation c/o DESY, Notkestraße 85, 22603, Hamburg, Germany

    Cy M. Jeffries & Dmitri I. Svergun

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  1. Cy M. Jeffries
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  2. Dmitri I. Svergun
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Correspondence to Dmitri I. Svergun .

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  1. Oxford Protein Production Facility UK, Didcot, Oxfordshire, United Kingdom

    Raymond J. Owens

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Jeffries, C.M., Svergun, D.I. (2015). High-Throughput Studies of Protein Shapes and Interactions by Synchrotron Small-Angle X-Ray Scattering. In: Owens, R. (eds) Structural Proteomics. Methods in Molecular Biology, vol 1261. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2230-7_15

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  • DOI: https://doi.org/10.1007/978-1-4939-2230-7_15

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