A1 Vertaisarvioitu alkuperäisartikkeli tieteellisessä lehdessä
Production and preliminary analysis of perdeuterated yeast inorganic pyrophosphatase crystals suitable for neutron diffraction
Tekijät: Tuominen VU, Myles DAA, Dauvergne MT, Lahti R, Heikinheimo P, Goldman A
Kustantaja: WILEY-BLACKWELL
Julkaisuvuosi: 2004
Journal: Acta Crystallographica Section D: Biological Crystallography
Tietokannassa oleva lehden nimi: ACTA CRYSTALLOGRAPHICA SECTION D-BIOLOGICAL CRYSTALLOGRAPHY
Lehden akronyymi: ACTA CRYSTALLOGR D
Vuosikerta: 60
Aloitussivu: 606
Lopetussivu: 609
Sivujen määrä: 4
ISSN: 0907-4449
DOI: https://doi.org/10.1107/S0907444903029585
Tiivistelmä
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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