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

Midori Taketa, Hanae Nakagawa, Mao Habukawa, Hisao Osuka, Kiyohito Kihira, Hirofumi Komori, Naoki Shibata, Masaharu Ishii, Yasuo Igarashi, Hirofumi Nishihara, Ki Seok Yoon, Seiji Ogo, Yasuhito Shomura, Yoshiki Higuchi

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    1 Citation (Scopus)


    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 2.58Å resolution and belonged to space group C2, with unit-cell parameters a = 131.43, b = 189.71, c = 124.59Å, β = 109.42°. Assuming the presence of two NAD+-reducing [NiFe] hydrogenase molecules in the asymmetric unit, V M was calculated to be 2.2Å3 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.

    Original languageEnglish
    Pages (from-to)96-99
    Number of pages4
    JournalActa Crystallographica Section F:Structural Biology Communications
    Publication statusPublished - Jan 1 2015

    All Science Journal Classification (ASJC) codes

    • Biophysics
    • Structural Biology
    • Biochemistry
    • Genetics
    • Condensed Matter Physics


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