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X-ray scattering experiments with high-flux X-ray source coupled rapid mixing microchannel device and their potential for high-flux neutron scattering investigations

The European Physical Journal E volume 36, Article number: 109 (2013) Cite this article

Abstract

Small-angle X-ray scattering provides global, shape-sensitive structural information about macromolecules in solution. Its extension to time dimension in the form of time-resolved SAXS investigations and combination with other time-resolved biophysical methods contributes immensely to the study of protein dynamics. TR-SAXS can also provide unique information about the global structures of transient intermediates during protein dynamics. An experimental set-up with low protein consumption is essential for an extensive use of TR-SAXS experiments on protein dynamics. In this direction, a newly developed 20-microchannel microfluidic continuous-flow mixer was combined with SAXS. With this set-up, we demonstrate ubiquitin unfolding dynamics after rapid mixing with the chaotropic agent Guanidinium-HCl within milliseconds using only ∼ 40 nanoliters of the protein sample per scattering image. It is suggested that, in the future, this new TR-SAXS platform will help to increase the use of time-resolved small-angle X-ray scattering, wide-angle X-ray scattering and neutron scattering experiments for studying protein dynamics in the early millisecond regime. The potential research field for this set-up includes protein folding, protein misfolding, aggregation in amyloidogenic diseases, function of intrinsically disordered proteins and various protein-ligand interactions.

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Authors and Affiliations

  1. Structural Dynamics of (Bio)chemical Systems, MPI-BPC, Am Fassberg 11, 37077, Goettingen, Germany

    R. Jain, M. Petri, S. Sonnenkalb & S. Techert

  2. Biological Micro- and Nanotechnology, MPI-BPC, Am Fassberg 11, 37077, Goettingen, Germany

    S. Kirschbaum & T. P. Burg

  3. Center of Advanced European Studies and Research (caesar), Micro Systems Technology (MST), 53175, Bonn, Germany

    H. Feindt & S. Steltenkamp

  4. NMR-based Structural Biology, MPI-BPC, Am Fassberg 11, 37077, Goettingen, Germany

    S. Becker & C. Griesinger

  5. Paul Scherrer Institut, 5232, Villigen, Switzerland

    A. Menzel

  6. Structural Dynamics of (Bio)chemical Systems at the Photon Research Division, DESY, Notkesstrasse 85, 22607, Hamburg, Germany

    S. Techert

Authors
  1. R. Jain
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  2. M. Petri
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  3. S. Kirschbaum
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  4. H. Feindt
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  5. S. Steltenkamp
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  6. S. Sonnenkalb
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  7. S. Becker
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  8. C. Griesinger
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  9. A. Menzel
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  10. T. P. Burg
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  11. S. Techert
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Corresponding author

Correspondence to T. P. Burg.

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Jain, R., Petri, M., Kirschbaum, S. et al. X-ray scattering experiments with high-flux X-ray source coupled rapid mixing microchannel device and their potential for high-flux neutron scattering investigations. Eur. Phys. J. E 36, 109 (2013). https://doi.org/10.1140/epje/i2013-13109-9

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  • DOI: https://doi.org/10.1140/epje/i2013-13109-9

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