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A device for the high-pressure oxygenation of protein crystals

A device for the high-pressure oxygenation of protein crystals
A device for the high-pressure oxygenation of protein crystals
A system has been developed for subjecting protein crystals to hyperbaric pressures of oxygen gas in order to promote enzymatic reaction. Crystals of an oxygenase or oxidase enzyme are grown anaerobically by hanging drop vapor diffusion, under crystallization conditions modified to eliminate combustible materials such as plastic coverslips and grease. The crystalline enzyme: substrate complex can then be exposed to oxygen gas at pressures up to 60 bar using a custom-built device or "bomb." In this way, reaction is initiated synchronously throughout the crystal and subsequent flash freezing allows the trapping of enzyme:product complexes in high occupancy. These complexes can then be structurally characterized by conventional monochromatic X-ray crystallography. The bomb is furnished from naval brass and lubricated with Fomblin RT15 perfluorinated polyether grease in order to ensure compatibility with the highly oxidizing environment.
protein crystallography, time-resolved studies, enzyme intermediates, high-pressure oxygen, nonheme iron oxygenaseisomorphous xenon derivatives, cryocrystallography
0003-2697
265-268
Rutledge, Peter J.
1bb10fcb-ce8b-4f02-8fe0-756ded021a40
Burzlaff, Nicolai I.
6a717e4c-c9aa-4b95-8170-73c99b01e2fd
Elkins, Jonathan M.
103a8855-1ca0-4abb-9ce2-a7c4f350fe39
Pickford, Michael
1cfc4137-cec7-41d2-9080-c2ab69ce5c64
Baldwin, Jack E.
8ee0f163-e32e-4e77-bf41-a2de55778ef6
Roach, Peter L.
ca94060c-4443-482b-af3e-979243488ba9
Rutledge, Peter J.
1bb10fcb-ce8b-4f02-8fe0-756ded021a40
Burzlaff, Nicolai I.
6a717e4c-c9aa-4b95-8170-73c99b01e2fd
Elkins, Jonathan M.
103a8855-1ca0-4abb-9ce2-a7c4f350fe39
Pickford, Michael
1cfc4137-cec7-41d2-9080-c2ab69ce5c64
Baldwin, Jack E.
8ee0f163-e32e-4e77-bf41-a2de55778ef6
Roach, Peter L.
ca94060c-4443-482b-af3e-979243488ba9

Rutledge, Peter J., Burzlaff, Nicolai I., Elkins, Jonathan M., Pickford, Michael, Baldwin, Jack E. and Roach, Peter L. (2002) A device for the high-pressure oxygenation of protein crystals. Analytical Biochemistry, 308 (2), 265-268. (doi:10.1016/S0003-2697(02)00246-4).

Record type: Article

Abstract

A system has been developed for subjecting protein crystals to hyperbaric pressures of oxygen gas in order to promote enzymatic reaction. Crystals of an oxygenase or oxidase enzyme are grown anaerobically by hanging drop vapor diffusion, under crystallization conditions modified to eliminate combustible materials such as plastic coverslips and grease. The crystalline enzyme: substrate complex can then be exposed to oxygen gas at pressures up to 60 bar using a custom-built device or "bomb." In this way, reaction is initiated synchronously throughout the crystal and subsequent flash freezing allows the trapping of enzyme:product complexes in high occupancy. These complexes can then be structurally characterized by conventional monochromatic X-ray crystallography. The bomb is furnished from naval brass and lubricated with Fomblin RT15 perfluorinated polyether grease in order to ensure compatibility with the highly oxidizing environment.

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More information

Published date: 15 September 2002
Keywords: protein crystallography, time-resolved studies, enzyme intermediates, high-pressure oxygen, nonheme iron oxygenaseisomorphous xenon derivatives, cryocrystallography

Identifiers

Local EPrints ID: 19846
URI: http://eprints.soton.ac.uk/id/eprint/19846
ISSN: 0003-2697
PURE UUID: 041555a3-005e-4500-bf52-ff2478badbf4
ORCID for Peter L. Roach: ORCID iD orcid.org/0000-0001-9880-2877

Catalogue record

Date deposited: 21 Feb 2006
Last modified: 08 Jan 2022 06:46

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Contributors

Author: Peter J. Rutledge
Author: Nicolai I. Burzlaff
Author: Jonathan M. Elkins
Author: Michael Pickford
Author: Jack E. Baldwin
Author: Peter L. Roach ORCID iD

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