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

Research output: Contribution to journalArticle

1 Citation (Scopus)

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

Fingerprint

hydrogen dehydrogenase
X ray analysis
Crystallization
NAD
X-Rays
crystallization
inoculation
purification
crystals
glycols
polyethylenes
isolation
x rays
vapors
cells
Cloud seeding
wavelengths
atoms
molecules
Crystals

All Science Journal Classification (ASJC) codes

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

Cite this

Crystallization and preliminary X-ray analysis of the NAD+-reducing [NiFe] hydrogenase from Hydrogenophilus thermoluteolus TH-1. / Taketa, Midori; Nakagawa, Hanae; Habukawa, Mao; Osuka, Hisao; Kihira, Kiyohito; Komori, Hirofumi; Shibata, Naoki; Ishii, Masaharu; Igarashi, Yasuo; Nishihara, Hirofumi; Yoon, Ki Seok; Ogo, Seiji; Shomura, Yasuhito; Higuchi, Yoshiki.

In: Acta Crystallographica Section F:Structural Biology Communications, Vol. 71, 01.01.2015, p. 96-99.

Research output: Contribution to journalArticle

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 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, vol. 71, pp. 96-99. https://doi.org/10.1107/S2053230X14026521
Taketa, Midori ; Nakagawa, Hanae ; Habukawa, Mao ; Osuka, Hisao ; Kihira, Kiyohito ; Komori, Hirofumi ; Shibata, Naoki ; Ishii, Masaharu ; Igarashi, Yasuo ; Nishihara, Hirofumi ; Yoon, Ki Seok ; Ogo, Seiji ; Shomura, Yasuhito ; Higuchi, Yoshiki. / Crystallization and preliminary X-ray analysis of the NAD+-reducing [NiFe] hydrogenase from Hydrogenophilus thermoluteolus TH-1. In: Acta Crystallographica Section F:Structural Biology Communications. 2015 ; Vol. 71. pp. 96-99.
@article{7fa5ab5875dc4bf9ae4cba622652dd4f,
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{\AA} resolution and belonged to space group C2, with unit-cell parameters a = 131.43, b = 189.71, c = 124.59{\AA}, 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{\AA}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.",
author = "Midori Taketa and Hanae Nakagawa and Mao Habukawa and Hisao Osuka and Kiyohito Kihira and Hirofumi Komori and Naoki Shibata and Masaharu Ishii and Yasuo Igarashi and Hirofumi Nishihara and Yoon, {Ki Seok} and Seiji Ogo and Yasuhito Shomura and Yoshiki Higuchi",
year = "2015",
month = "1",
day = "1",
doi = "10.1107/S2053230X14026521",
language = "English",
volume = "71",
pages = "96--99",
journal = "Acta Crystallographica Section F:Structural Biology Communications",
issn = "1744-3091",
publisher = "John Wiley and Sons Ltd",

}

TY - JOUR

T1 - Crystallization and preliminary X-ray analysis of the NAD+-reducing [NiFe] hydrogenase from Hydrogenophilus thermoluteolus TH-1

AU - Taketa, Midori

AU - Nakagawa, Hanae

AU - Habukawa, Mao

AU - Osuka, Hisao

AU - Kihira, Kiyohito

AU - Komori, Hirofumi

AU - Shibata, Naoki

AU - Ishii, Masaharu

AU - Igarashi, Yasuo

AU - Nishihara, Hirofumi

AU - Yoon, Ki Seok

AU - Ogo, Seiji

AU - Shomura, Yasuhito

AU - Higuchi, Yoshiki

PY - 2015/1/1

Y1 - 2015/1/1

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=84921747647&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84921747647&partnerID=8YFLogxK

U2 - 10.1107/S2053230X14026521

DO - 10.1107/S2053230X14026521

M3 - Article

C2 - 25615977

AN - SCOPUS:84921747647

VL - 71

SP - 96

EP - 99

JO - Acta Crystallographica Section F:Structural Biology Communications

JF - Acta Crystallographica Section F:Structural Biology Communications

SN - 1744-3091

ER -