Abstract
In many biomolecular interactions, changes in the assembly states and structural conformations of participants can act as a complementary reporter of binding to functional and thermodynamic assays. This structural information is captured by a number of structural biology and biophysical techniques that are viable either as primary screens in small-scale applications or as secondary screens to complement higher throughput methods. In particular, small-angle X-ray scattering (SAXS) reports the average distance distribution between all atoms after orientational averaging. Such information is important when for example investigating conformational changes involved in inhibitory and regulatory mechanisms where binding events do not necessarily cause functional changes. Thus, we summarise here the current and prospective capabilities of SAXS-based screening in the context of other methods that yield structural information. Broad guidelines are also provided to assist readers in preparing screening protocols that are tailored to available X-ray sources.
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Notes
Based on observed droplet volume of order 20 nl and \(\sim 100\) droplets required to produce the reported average using 100 ms exposure time at BM29, ESRF Grenoble.
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P.C. was supported by the EIPOD postdoctoral programme cofunded by the European Molecular Biology Laboratory (EMBL) and the Marie Curie Actions Cofund grant MSCACOFUND-FP no. 664726. J.H. gratefully acknowledges the EMBL and the German Research Council (Deutsche Forschungsgemeinschaft, DFG) for support via an Emmy-Noether Fellowship and the Priority Programme SPP1935.
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Chen, Pc., Hennig, J. The role of small-angle scattering in structure-based screening applications. Biophys Rev 10, 1295–1310 (2018). https://doi.org/10.1007/s12551-018-0464-x
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DOI: https://doi.org/10.1007/s12551-018-0464-x