Production and preliminary analysis of perdeuterated yeast inorganic pyrophosphatase crystals suitable for neutron diffraction

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 ord...

Ausführliche Beschreibung

Gespeichert in:
Bibliographische Detailangaben
Veröffentlicht in:Acta crystallographica. Section D, Biological crystallography. Biological crystallography., 2004-03, Vol.60 (3), p.606-609
Hauptverfasser: Tuominen, Vesa U., Myles, Dean A. A., Dauvergne, Marie-Thérèse, Lahti, Reijo, Heikinheimo, Pirkko, Goldman, Adrian
Format: Artikel
Sprache:eng
Schlagworte:
Online-Zugang:Volltext
Tags: Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
Beschreibung
Zusammenfassung: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.
ISSN:1399-0047
0907-4449
1399-0047
DOI:10.1107/S0907444903029585