Production and preliminary analysis of perdeuterated yeast inorganic pyrophosphatase crystals suitable for neutron diffraction.
作者： Vesa U. Tuominen , Dean A. A. Myles , Marie-Thérèse Dauvergne , Reijo Lahti , Pirkko Heikinheimo
DOI: 10.1107/S0907444903029585
关键词:
摘要: 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 positions protons in active site, which can be best achieved neutron diffraction analysis. In order to reduce hydrogen incoherent-scattering background and improve signal-to-noise ratio reflections, deuterated protein was produced. Deuterated 96% enriched with deuterium produced high yield crystals as large 2 mm on one side were obtained. These have unit-cell parameters = 58.9, b 103.9, c 117.0 A, alpha beta gamma 90 degrees at 273 K diffract neutrons resolutions 2.5-3 A. The X-ray structure perdeuterated has also been refined 1.9 A resolution.
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