Recombinant formate dehydrogenase from Arabidopsis thaliana : Preparation, crystal growth in microgravity, and prelimina
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Recombinant Formate Dehydrogenase from Arabidopsis thaliana: Preparation, Crystal Growth in Microgravity, and Preliminary XRay Diffraction Study I. G. Shabalina,c, A. E. Serovb, O. E. Skirgellob, V. I. Timofeeva, V. R. Samyginaa, V. O. Popovc, V. I. Tishkovb, and I. P. Kuranovaa a
Shubnikov Institute of Crystallography, Russian Academy of Sciences, Leninskiі pr. 59, Moscow, 119333 Russia email: [email protected] b Faculty of Chemistry, Moscow State University, Moscow, 119992 Russia c Bach Institute of Biochemistry, Russian Academy of Sciences, Leninskiі pr. 33, Moscow, 119071 Russia Received March 31, 2010
Abstract—Crystals of highpurity recombinant NAD+dependent formate dehydrogenase from the higher plant Arabidopsis thaliana (AraFDH) were grown in microgravity in the Modul’1 protein crystallization apparatus on the International Space Station. The spacegrown crystals have larger sizes than those grown on Earth. Xray diffraction data suitable for determining the threedimensional structure were collected from the spacegrown crystals to a resolution of 1.22 Å using an Xray synchrotron source. The crystals belong to sp. gr. P43212; the unitcell parameters are a = b = 107.865 Å, c = 71.180 Å, α = β = γ = 90°. DOI: 10.1134/S1063774510050159
INTRODUCTION The growth of highquality protein crystals suitable for determining the threedimensional structures at high resolution is an important problem. The crystal lization in microgravity is one efficient approach to improving the Xray diffraction quality of crystals. The quality of the protein crystals depends significantly on the mass transfer to growing crystals, which occurs through convection or diffusion. In microgravity, where the convection is virtually absent, the diffusion transport dominates. A stable concentration gradient is established in the solution around the crystals so that the crystal growth occurs in the most favorable condi tions at a low supersaturation. Due to the absence of sedimentation, crystals do not fall to the bottom of the vessel, which yields the uniform transfer of protein molecules to all growing faces and facilitates the for mation of more isometric crystals in microgravity [1]. In 2006–2009 a series of experiments on the pro tein crystal growth in microgravity were carried out on the Russian Segment of the International Space Sta tion (RS ISS) [2]. Crystals of several proteins (includ ing geneticallyengineered human insulin, carbox ypeptidase, and sialylated insulin) were grown in the Modul’1 protein crystallization apparatus con structed in the Special Design Bureau of the Shubni kov Institute of Crystallography of the Russian Acad emy of Sciences. These crystals had larger sizes and were of a higher Xray diffraction quality than the earthgrown crystals of the corresponding proteins [2–4].
In this study we report the results of the crystalliza tion of recombinant NAD+dependent formate dehy drogenase from Arabidopsis thaliana (AraFDH) in the presence of sodium azide in th
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