Purification, crystallization and preliminary X-ray analysis of inositol dehydrogenase (IDH) from Bacillus subtilis.

Publication Type:

Journal Article

Authors:

Van Straaten, K E; Hoffort, A; Palmer, D R J; Sanders, D A R

Source:

Acta Crystallogr Sect F Struct Biol Cryst Commun, Volume 64, Issue Pt 2, p.98-101 (2008)

Keywords:

Bacillus subtilis, Catalysis, Crystallization, Crystallography, X-Ray, Electrophoresis, Polyacrylamide Gel, Kinetics, Oxidoreductases, Soil Microbiology, Spectrophotometry, Ultraviolet

Abstract:

<p>Inositol dehydrogenase (IDH) is an enzyme that catalyses the NAD(+)-dependent oxidation of myo-inositol to scyllo-inosose. The enzyme has been purified to homogeneity by means of Ni(2+)-affinity chromatography and was crystallized in both native and selenomethionine (SeMet) labelled forms using the microbatch method. SAD X-ray diffraction data were collected to 2.0 A resolution from a SeMet-labelled crystal at the Advanced Photon Source (APS) and a MAD data set was collected to 1.75 A resolution at the Canadian Light Source (CLS); this is the first reported anomalous diffraction experiment from the CLS. The crystals belong to space group I222 and contain one molecule per asymmetric unit.</p>