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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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 2.58Å 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
Volume71
DOIs
Publication statusPublished - Jan 1 2015

All Science Journal Classification (ASJC) codes

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

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    Taketa, M., Nakagawa, H., Habukawa, M., Osuka, H., Kihira, K., Komori, H., Shibata, N., Ishii, M., Igarashi, Y., Nishihara, H., Yoon, K. S., Ogo, S., Shomura, Y., & Higuchi, Y. (2015). Crystallization and preliminary X-ray analysis of the NAD+-reducing [NiFe] hydrogenase from Hydrogenophilus thermoluteolus TH-1. Acta Crystallographica Section F:Structural Biology Communications, 71, 96-99. https://doi.org/10.1107/S2053230X14026521