TY - JOUR
T1 - Production and preliminary analysis of perdeuterated yeast inorganic pyrophosphatase crystals suitable for neutron diffraction
AU - Tuominen, Vesa U.
AU - Myles, Dean A.A.
AU - Dauvergne, Marie Thérèse
AU - Lahti, Reijo
AU - Heikinheimo, Pirkko
AU - Goldman, Adrian
PY - 2004/3
Y1 - 2004/3
N2 - 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 Å, α = β = γ = 90° at 273 K and diffract neutrons to resolutions of 2.5-3 Å. The X-ray structure of the perdeuterated protein has also been refined at 273 K to 1.9 Å resolution.
AB - 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 Å, α = β = γ = 90° at 273 K and diffract neutrons to resolutions of 2.5-3 Å. The X-ray structure of the perdeuterated protein has also been refined at 273 K to 1.9 Å resolution.
UR - http://www.scopus.com/inward/record.url?scp=7444237598&partnerID=8YFLogxK
U2 - 10.1107/S0907444903029585
DO - 10.1107/S0907444903029585
M3 - Article
C2 - 14993708
AN - SCOPUS:7444237598
SN - 0907-4449
VL - 60
SP - 606
EP - 609
JO - Acta Crystallographica Section D: Biological Crystallography
JF - Acta Crystallographica Section D: Biological Crystallography
IS - 3
ER -