title: Crystallization and preliminary X-ray analysis of the NAD+-reducing [NiFe] hydrogenase from Hydrogenophilus thermoluteolus TH-1.
abstract: 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, β=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.
authors.affiliation: Department of Picobiology, University of Hyogo, 3-2-1 Koto, Kamigori-cho, Ako-gun, Hyogo 678-1297, Japan.
authors.forename: Midori
authors.initials: M
authors.lastname: Taketa
authors.affiliation: Department of Picobiology, University of Hyogo, 3-2-1 Koto, Kamigori-cho, Ako-gun, Hyogo 678-1297, Japan.
authors.forename: Hanae
authors.initials: H
authors.lastname: Nakagawa
authors.affiliation: Department of Picobiology, University of Hyogo, 3-2-1 Koto, Kamigori-cho, Ako-gun, Hyogo 678-1297, Japan.
authors.forename: Mao
authors.initials: M
authors.lastname: Habukawa
authors.affiliation: Department of Picobiology, University of Hyogo, 3-2-1 Koto, Kamigori-cho, Ako-gun, Hyogo 678-1297, Japan.
authors.forename: Hisao
authors.initials: H
authors.lastname: Osuka
authors.affiliation: Department of Picobiology, University of Hyogo, 3-2-1 Koto, Kamigori-cho, Ako-gun, Hyogo 678-1297, Japan.
authors.forename: Kiyohito
authors.initials: K
authors.lastname: Kihira
authors.affiliation: Department of Picobiology, University of Hyogo, 3-2-1 Koto, Kamigori-cho, Ako-gun, Hyogo 678-1297, Japan.
authors.forename: Hirofumi
authors.initials: H
authors.lastname: Komori
authors.affiliation: Department of Picobiology, University of Hyogo, 3-2-1 Koto, Kamigori-cho, Ako-gun, Hyogo 678-1297, Japan.
authors.forename: Naoki
authors.initials: N
authors.lastname: Shibata
authors.affiliation: Department of Biotechnology, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan.
authors.forename: Masaharu
authors.initials: M
authors.lastname: Ishii
authors.affiliation: Department of Biotechnology, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan.
authors.forename: Yasuo
authors.initials: Y
authors.lastname: Igarashi
authors.affiliation: Department of Bioresource Science, Ibaraki University, 3-21-1 Chu-ou, Ami, Ibaraki 300-0393, Japan.
authors.forename: Hirofumi
authors.initials: H
authors.lastname: Nishihara
authors.affiliation: International Institute for Carbon Neutral Energy Research (WPI-I2CNER), Kyushu University, 744 Moto-oka, Nishi-ku, Fukuoka 819-0395, Japan.
authors.forename: Ki-Seok
authors.initials: KS
authors.lastname: Yoon
authors.affiliation: International Institute for Carbon Neutral Energy Research (WPI-I2CNER), Kyushu University, 744 Moto-oka, Nishi-ku, Fukuoka 819-0395, Japan.
authors.forename: Seiji
authors.initials: S
authors.lastname: Ogo
authors.affiliation: Department of Picobiology, University of Hyogo, 3-2-1 Koto, Kamigori-cho, Ako-gun, Hyogo 678-1297, Japan.
authors.forename: Yasuhito
authors.initials: Y
authors.lastname: Shomura
authors.affiliation: Department of Picobiology, University of Hyogo, 3-2-1 Koto, Kamigori-cho, Ako-gun, Hyogo 678-1297, Japan.
authors.forename: Yoshiki
authors.initials: Y
authors.lastname: Higuchi
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry*; D003460:Crystallization; D018360:Crystallography, X-Ray; D006864:Hydrogenase / Q000737:chemistry*; D043363:Hydrogenophilaceae / Q000201:enzymology*
chemical_list: D001426:Bacterial Proteins; C043296:nickel-iron hydrogenase; D006864:Hydrogenase
keywords: NAD+-reducing [NiFe] hydrogenase; diaphorase; hydrogen metabolism; respiratory complex I