A1 Refereed original research article in a scientific journal
Production and preliminary analysis of perdeuterated yeast inorganic pyrophosphatase crystals suitable for neutron diffraction
Authors: Tuominen VU, Myles DAA, Dauvergne MT, Lahti R, Heikinheimo P, Goldman A
Publisher: WILEY-BLACKWELL
Publication year: 2004
Journal: Acta Crystallographica Section D: Biological Crystallography
Journal name in source: ACTA CRYSTALLOGRAPHICA SECTION D-BIOLOGICAL CRYSTALLOGRAPHY
Journal acronym: ACTA CRYSTALLOGR D
Volume: 60
First page : 606
Last page: 609
Number of pages: 4
ISSN: 0907-4449
DOI: https://doi.org/10.1107/S0907444903029585
Abstract
Yeast inorganic pyrophosphatase (Y-PPase) is a model system for studying phosphoryl-transfer reactions catalysed by multiple metal ions. To understand the process requires knowledge of the positions of the protons in the active site, which can be best achieved by neutron diffraction analysis. In order to reduce the hydrogen incoherent-scattering background and to improve the signal-to-noise ratio of the neutron reflections, deuterated protein was produced. Deuterated protein 96% enriched with deuterium was produced in high yield and crystals as large as 2 mm on one side were obtained. These crystals have unit-cell parameters a=58.9, b=103.9, c=117.0 Angstrom, alpha=beta=gamma=90degrees at 273 K and diffract neutrons to resolutions of 2.5-3 Angstrom. The X-ray structure of the perdeuterated protein has also been refined at 273 K to 1.9 Angstrom resolution.
Yeast inorganic pyrophosphatase (Y-PPase) is a model system for studying phosphoryl-transfer reactions catalysed by multiple metal ions. To understand the process requires knowledge of the positions of the protons in the active site, which can be best achieved by neutron diffraction analysis. In order to reduce the hydrogen incoherent-scattering background and to improve the signal-to-noise ratio of the neutron reflections, deuterated protein was produced. Deuterated protein 96% enriched with deuterium was produced in high yield and crystals as large as 2 mm on one side were obtained. These crystals have unit-cell parameters a=58.9, b=103.9, c=117.0 Angstrom, alpha=beta=gamma=90degrees at 273 K and diffract neutrons to resolutions of 2.5-3 Angstrom. The X-ray structure of the perdeuterated protein has also been refined at 273 K to 1.9 Angstrom resolution.
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