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Crystallization and preliminary X‐ray analysis of the NAD + ‐reducing [NiFe] hydrogenase from Hydrogenophilus thermoluteolus TH‐1
Author(s) -
Taketa Midori,
Nakagawa Hanae,
Habukawa Mao,
Osuka Hisao,
Kihira Kiyohito,
Komori Hirofumi,
Shibata Naoki,
Ishii Masaharu,
Igarashi Yasuo,
Nishihara Hirofumi,
Yoon KiSeok,
Ogo Seiji,
Shomura Yasuhito,
Higuchi Yoshiki
Publication year - 2015
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 37
ISSN - 2053-230X
DOI - 10.1107/s2053230x14026521
Subject(s) - crystallization , hydrogenase , nad+ kinase , materials science , x ray , nuclear chemistry , chemistry , crystallography , catalysis , physics , nuclear physics , enzyme , biochemistry , organic chemistry
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 C 2, 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, V M was calculated to be 2.2 Å 3 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.