Online Raman spectroscopy for structural biology on beamline ID29 of the ESRF

J Struct Biol. 2017 Nov;200(2):124-127. doi: 10.1016/j.jsb.2017.10.004. Epub 2017 Oct 16.

Abstract

Raman spectroscopy can probe the structure and conformations of specific chemical groups within proteins and may thus be used as a technique complementary to X-ray crystallography. This combined approach can be decisive in resolving ambiguities in the interpretation of enzymatic or X-ray induced processes. Here, we present an online Raman setup developed at the European Synchrotron that allows for interleaved Raman spectra acquisition and X-ray diffraction measurements with fast probe exchange and simple alignment while maintaining a high sensitivity over the entire spectral range. This device has been recently employed in the study of a covalent intermediate in the O2-dependent breakdown of uric acid by the cofactor-free enzyme urate oxidase and to monitor its decay induced by X-ray exposure.

Keywords: Diffraction-complementary technique; Kinetic crystallography; Macromolecular crystallography; Radiation damage; Raman spectroscopy.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Crystallography, X-Ray / methods
  • Molecular Conformation
  • Spectrum Analysis, Raman / methods*
  • Synchrotrons
  • Urate Oxidase / metabolism*
  • Uric Acid / analogs & derivatives
  • Uric Acid / chemistry*
  • X-Ray Diffraction / methods

Substances

  • 5-hydroxyisourate
  • Uric Acid
  • Urate Oxidase