title: A device for the high-pressure oxygenation of protein crystals.
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.
authors.affiliation: The Dyson Perrins Laboratory and the Oxford Centre for Molecular Sciences, University of Oxford, UK. peter.rutledge@chem.ox.ac.uk
authors.forename: Peter J
authors.initials: PJ
authors.lastname: Rutledge
authors.forename: Nicolai I
authors.initials: NI
authors.lastname: Burzlaff
authors.forename: Jonathan M
authors.initials: JM
authors.lastname: Elkins
authors.forename: Michael
authors.initials: M
authors.lastname: Pickford
authors.forename: Jack E
authors.initials: JE
authors.lastname: Baldwin
authors.forename: Peter L
authors.initials: PL
authors.lastname: Roach
mesh_terms: D000818:Animals; D003460:Crystallization; D018360:Crystallography, X-Ray; D005296:Ferrous Compounds / Q000378:metabolism*; D006801:Humans; D010088:Oxidoreductases / Q000378:metabolism*; D010100:Oxygen / Q000378:metabolism*; D010106:Oxygenators
chemical_list: D005296:Ferrous Compounds; D010088:Oxidoreductases; C038370:isopenicillin N synthetase; D010100:Oxygen