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Mix-and-Inject Serial Femtosecond Crystallography to Capture RNA Riboswitch Intermediates

  1. Author:
    Stagno,Jason
    Knoska, Juraj
    Chapman, Henry N
    Wang,Yun-Xing

  2. Author Address

    Protein-Nucleic Acid Interaction Section, Center for Structural Biology, Center for Cancer Research, National Cancer Institute, Frederick, MD, USA., Center for Free-Electron Laser Science, Deutsches Elektronen-Synchrotron DESY, Hamburg, Germany., Department of Physics, Universit 228;t Hamburg, Hamburg, Germany., Centre for Ultrafast Imaging, Universit 228;t Hamburg, Hamburg, Germany., Protein-Nucleic Acid Interaction Section, Center for Structural Biology, Center for Cancer Research, National Cancer Institute, Frederick, MD, USA. wangyunx@mail.nih.gov.,
    1. Year: 2023
  2. Journal: Methods in Molecular Biology (Clifton, N.J.)
    1. 2568
    2. Pages: 243-249
  4. Type of Article: Article
  1. Abstract:

    Time-resolved structure determination of macromolecular conformations and ligand-bound intermediates is extremely challenging, particularly for RNA. With rapid technological advances in both microfluidic liquid injection and X-ray free electron lasers (XFEL), a new frontier has emerged in time-resolved crystallography whereby crystals can be mixed with ligand and then probed with X-rays (mix-and-inject) in real time and at room temperature. This chapter outlines the basic setup and procedures for mix-and-inject experiments for recording time-resolved crystallographic data of riboswitch RNA reaction states using serial femtosecond crystallography (SFX) and an XFEL. © 2023. This is a U.S. government work and not under copyright protection in the U.S.; foreign copyright protection may apply.

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  2. DOI: 10.1007/978-1-0716-2687-0_16
  3. PMID: 36227573

Library Notes

  1. Fiscal Year: FY2022-2023
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