Purification, crystallization and preliminary X-ray diffraction studies of a putative UDP-N-acetyl-D-mannosamine dehydrogenase from Pyrococcus horikoshii OT3

Neratur K Lokanath; Kudigana J Pampa; Toshimi Kamiya; Naoki Kunishima

doi:10.1107/S1744309107016685

Purification, crystallization and preliminary X-ray diffraction studies of a putative UDP-N-acetyl-D-mannosamine dehydrogenase from Pyrococcus horikoshii OT3

Acta Crystallogr Sect F Struct Biol Cryst Commun. 2007 May 1;63(Pt 5):412-4. doi: 10.1107/S1744309107016685. Epub 2007 Apr 14.

Authors

Neratur K Lokanath¹, Kudigana J Pampa, Toshimi Kamiya, Naoki Kunishima

Affiliation

¹ Advanced Protein Crystallography Research Group, RIKEN SPring-8 Center, Harima Institute, 1-1-1 Kouto, Sayo-cho, Sayo-gun, Hyogo 679-5148, Japan.

Abstract

A putative UDP-N-acetyl-D-mannosamine dehydrogenase from Pyrococcus horikoshii OT3, an essential enzyme for polysaccharide biosynthesis, has been overexpressed in Escherichia coli and purified. Crystals were obtained using the oil-microbatch method at 291 K. A native data set extending to 1.8 A resolution has been collected and processed in space group P2(1). Assuming the presence of a dimer in the asymmetric unit, the V(M) value is calculated to be 2.3 A3 Da(-1), which is consistent with the result of a dynamic light-scattering experiment that shows a dimeric state of the protein in solution.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Crystallization
Crystallography, X-Ray
Oxidoreductases / chemistry*
Protein Conformation
Pyrococcus horikoshii / enzymology*

Substances

Oxidoreductases