Search research articles

ABOUT JoVE

Overview Leadership Blog JoVE Help Center

AUTHORS

Publishing Process Editorial Board Scope & Policies Peer Review FAQ Submit

LIBRARIANS

Testimonials Subscriptions Access Resources Library Advisory Board FAQ

RESEARCH

JoVE Journal Methods Collections JoVE Encyclopedia of Experiments Archive

EDUCATION

JoVE Core JoVE Business JoVE Science Education JoVE Lab Manual Faculty Resource Center Faculty Site

Terms & Conditions of Use

Search research articles

Related Experiment Video

Updated: Dec 8, 2025

Fixed Target Serial Data Collection at Diamond Light Source

Fixed Target Serial Data Collection at Diamond Light Source

Published on: February 26, 2021

Anaerobic fixed-target serial crystallography.

Patrick Rabe¹, John H Beale², Agata Butryn²

¹Chemistry Research Laboratory, University of Oxford, 12 Mansfield Road, Oxford OX1 3TA, United Kingdom.

|September 17, 2020

Summary

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by

Same authorSame journal

Towards light-coupled sample preparation for time-resolved cryoEM studies.

IUCrJ·2026

Same author

Correction to "Monooxygenase Activity of Indoleamine 2,3-Dioxygenase".

Journal of the American Chemical Society·2026

Same author

Illuminating the molecular basis of human daylight vision.

Science (New York, N.Y.)·2026

Same author

Structure of the pre-initiation complex explains CMGE biogenesis.

Nature·2026

Same author

Substrate-Selective Inhibition of the SARS-CoV-2 Papain-Like Protease: Inhibition of Hydrolysis of Human Over Viral Substrates.

Chemistry (Weinheim an der Bergstrasse, Germany)·2026

Same author

Drop-on-fixed-target reaction initiation approach for serial and time-resolved crystallography.

IUCrJ·2026

Same journal

Cryo-EM analysis of cooperative conformational changes in the SARS-CoV-2 spike protein trimer.

IUCrJ·2026

Same journal

Towards time-resolved MicroED grid preparation using mix-and-inject gas dynamic virtual nozzles.

IUCrJ·2026

Same journal

How cryoEM has advanced our understanding of bacteriophages and bacteriocins targeting Clostridioides difficile.

IUCrJ·2026

Same journal

CryoEM structures reveal allosteric regulation of the catalytic activity of the multi-protein human MAT enzyme complexes.

IUCrJ·2026

Same journal

Cryo-EM-guided subtractive optimization of a novel VCP/p97 inhibitor.

IUCrJ·2026

See all related articles

This summary is machine-generated.

This study introduces a new method for room-temperature X-ray diffraction data collection of oxygen-sensitive enzymes. The technique preserves anaerobic conditions, enabling studies on crucial enzymes like VioC, AlkB, and IPNS.

Area of Science:

Biochemistry
Structural Biology
Crystallography

Background:

Cryo-X-ray diffraction is standard for oxygen-sensitive enzymes, but limits reactivity.
Room-temperature serial methods offer advantages but lack anaerobic conditions for dependent enzymes.
Previous methods have not explored maintaining anaerobic environments for room-temperature serial data collection at synchrotrons.

Purpose of the Study:

To develop and validate a methodology for room-temperature serial X-ray diffraction data collection of anaerobic samples.
To enable studies on oxygen-dependent enzymes that are sensitive to X-ray radiation.
To adapt existing sample mounting techniques for low-dose, room-temperature, anaerobic data collection.

Main Methods:

Adaptation of the 'sheet-on-sheet' sample mount for anaerobic conditions.

Keywords:

2-oxoglutarate/α-ketoglutarate oxygenases anaerobic crystallization oxygen-employing enzymes penicillin biosynthesis serial crystallography

More Related Videos

$Microfluidic Chips for In Situ Crystal X-ray Diffraction and In Situ Dynamic Light Scattering for Serial Crystallography$

Microfluidic Chips for In Situ Crystal X-ray Diffraction and In Situ Dynamic Light Scattering for Serial Crystallography

Published on: April 24, 2018

$On-Chip Crystallization and Large-Scale Serial Diffraction at Room Temperature$

On-Chip Crystallization and Large-Scale Serial Diffraction at Room Temperature

Published on: March 11, 2022

Related Experiment Videos

Last Updated: Dec 8, 2025

Fixed Target Serial Data Collection at Diamond Light Source

Fixed Target Serial Data Collection at Diamond Light Source

Published on: February 26, 2021

$Microfluidic Chips for In Situ Crystal X-ray Diffraction and In Situ Dynamic Light Scattering for Serial Crystallography$

Microfluidic Chips for In Situ Crystal X-ray Diffraction and In Situ Dynamic Light Scattering for Serial Crystallography

Published on: April 24, 2018

$On-Chip Crystallization and Large-Scale Serial Diffraction at Room Temperature$

On-Chip Crystallization and Large-Scale Serial Diffraction at Room Temperature

Published on: March 11, 2022

Sample preparation in an anaerobic glovebox with gentle crystal handling.

Low-dose, room-temperature serial data collection at synchrotron light sources.

Main Results:

Successful methodology for preserving localized anaerobic environments (<5 min) during data collection.
Demonstrated utility with X-ray radiation-sensitive Fe(II)-containing enzymes: VioC, AlkB, and isopenicillin N synthase (IPNS).
Method shows low sample consumption and easy preparation.

Conclusions:

The developed method facilitates room-temperature serial X-ray diffraction of anaerobic, oxygen-sensitive enzymes.
This technique expands the scope of synchrotron-based crystallographic studies for critical enzyme classes.
Enables structural insights into enzymes involved in DNA repair and antibiotic biosynthesis.