Exploiting fourth-generation synchrotron radiation for enzyme and photoreceptor characterization

IUCrJ. 2025 Jan 1;12(Pt 1):36-48. doi: 10.1107/S2052252524010868.

Abstract

The upgrade of the European Synchrotron Radiation Facility (ESRF) in Grenoble, France to an Extremely Brilliant Source (EBS) is expected to enable time-resolved synchrotron serial crystallography (SSX) experiments with sub-millisecond time resolution. ID29 is a new beamline dedicated to SSX experiments at ESRF-EBS. Here, we report experiments emerging from the initial phase of user operation at ID29. We first used microcrystals of photoactive yellow protein as a model system to exploit the potential of microsecond pulses for SSX. Subsequently, we investigated microcrystals of cytochrome c nitrite reductase (ccNiR) with microsecond X-ray pulses. CcNiR is a decaheme protein that is ideal for the investigation of radiation damage at the various heme-iron sites. Finally, we performed a proof-of-concept subsecond time-resolved SSX experiment by photoactivating microcrystals of a myxobacterial phytochrome.

Keywords: environmental chemistry; macromolecular machines; radiation damage; structural biology; structure determination; time-resolved synchrotron serial crystallography.

MeSH terms

  • Bacterial Proteins* / chemistry
  • Bacterial Proteins* / metabolism
  • Crystallography, X-Ray / methods
  • Photoreceptors, Microbial* / chemistry
  • Photoreceptors, Microbial* / metabolism
  • Phytochrome / chemistry
  • Phytochrome / metabolism
  • Synchrotrons*

Substances

  • Bacterial Proteins
  • Photoreceptors, Microbial
  • photoactive yellow protein, Bacteria
  • Phytochrome