Completed Cells, Biochemistry & Physiology Chemistry

Dynamic Structural Biology: New Tools and Strategies for General Applications

Funder Wellcome Charity-funded
Recipient organisation Diamond Light Source Limited Source-published name: Diamond Light Source LTD
Funding £2.5M
Period Jun 2018 — May 2023

In plain English

AI plain-English summary

Structural biologists have spent decades freezing their samples to 100 Kelvin to get clear images, but a frozen enzyme cannot do its job—so researchers are now building tools to watch proteins work at room temperature instead. This matters because current methods like X-ray crystallography and cryo-EM capture only static snapshots of biomolecules, while life depends on dynamic movements and chemical reactions that happen in milliseconds. The team will develop new ways to mix tiny crystals with substrates, then fire ultrafast X-ray pulses from free-electron lasers and synchrotrons to record both the atomic structure and the electronic state of enzymes as they catalyse reactions. They focus on two enzyme families: one that forms a highly reactive iron-oxo intermediate crucial for many biological processes, and beta-lactamases, which break down antibiotics and drive antimicrobial resistance. If successful, the project will turn time-resolved structural biology from a specialist technique into a general tool. This could eventually help design better antibiotics that evade bacterial defences, or engineer industrial enzymes that work faster at ambient temperatures. The research is fundamental science—it asks how enzymes actually move and change during catalysis—but understanding those motions is the first step toward controlling them.

View original technical description
X-ray crystallography and cryo-EM are essential tools for structural biologists, but these data are almost always from samples held at 100 K. Life is dynamic and function is not compatible with the cryogenic conditions. X-ray Free Electron Lasers (XFELs) offer new opportunities because their unparalleled intensity reduces the crystal size requirements such that even submicron size crystals yield high quality structures.1-3 The fs pulse provides extraordinary temporal resolution and data without radiation-induced alterations.4-9 XFELs are motivating new serial data collection methods at room temperature.10-13 Synchrotron beamlines like Diamond’s VMXi with high-flux, micro-focus, and fast detector are also pushing serial methods that complement XFELs. What is critically missing are tools to manipulate microcrystal samples and to correlate enzyme kinetics, spectroscopy, and time-resolved structural biology. Therefore, the specific aims of this proposal are to i) extend and generalize experimental methods in enzyme kinetics to microcrystalline samples, and ii) and iii) to develop time-resolved structural biology in general at XFELs (ii) and at synchrotrons (iii). In particular, we will correlate room-temperature, serial crystallographic and spectroscopic data to determine the electronic and atomic structures of metalloenzymes engaged in catalysis. Some of our enzymes create Fe(IV)=O intermediates, whereas beta-lactamases are responsible for antimicrobial resistance.

View the original record at the funder ↗

Researchers

Allen Orville (EPMC Awardee)

Related Research

Grants with similar aims, by meaning.

Breaking the Cage: Transformative Time-resolved Crystallography using Fixed Targets at Synchrotrons and XFELs
Time-resolved methodologies to provide both spatial and temporal resolution in Electron Microscopy
Cryo-EM for understanding molecular processes in health and disease
Developing novel approaches for time resolved structural biology.
National Cryo-Electron Microscopy Facility

Original classification

Investigator Award in Science

Plain English summaries and category classifications on this site are generated by AI and may not perfectly reflect the original research.