Crystallization and preliminary X-ray analysis of the NAD+-reducing [NiFe] hydrogenase from Hydrogenophilus thermoluteolus TH-1.

Acta crystallographica. Section F, Structural biology communications

PubMedID: 25615977

Taketa M, Nakagawa H, Habukawa M, Osuka H, Kihira K, Komori H, Shibata N, Ishii M, Igarashi Y, Nishihara H, Yoon KS, Ogo S, Shomura Y, Higuchi Y. Crystallization and preliminary X-ray analysis of the NAD+-reducing [NiFe] hydrogenase from Hydrogenophilus thermoluteolus TH-1. Acta Crystallogr F Struct Biol Commun. 2015;71(Pt 1):96-9.
NAD+-reducing [NiFe] hydrogenases catalyze the oxidoreduction of dihydrogen concomitant with the interconversion of NAD+ and NADH. Here, the isolation, purification and crystallization of the NAD+-reducing [NiFe] hydrogenase from Hydrogenophilus thermoluteolus TH-1 are reported. Crystals of the NAD+-reducing [NiFe] hydrogenase were obtained within one week from a solution containing polyethylene glycol using the sitting-drop vapour-diffusion method and micro-seeding. The crystal diffracted to 2. 58 Å resolution and belonged to space group C2, with unit-cell parameters a=131. 43, b=189. 71, c=124. 59 Å, ß=109. 42°. Assuming the presence of two NAD+-reducing [NiFe] hydrogenase molecules in the asymmetric unit, VM was calculated to be 2. 2 Å3 Da(-1), which corresponds to a solvent content of 43%. Initial phases were determined by the single-wavelength anomalous dispersion method using the anomalous signal from the Fe atoms.