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Cloning, expression, purification, crystallization and preliminary crystallographic analysis of NifH2 from Methanocaldococcus jannaschii

Cloning, expression, purification, crystallization and preliminary crystallographic analysis of... Nitrogenases are protein complexes that are only found in Azotobacter and are required for biological nitrogen fixation. They are made up of a nitrogenase, which is a NifD2/NifK2 heterotetramer, and a nitrogenase reductase, which is a homodimer of NifH. Many homologues of nitrogenase have been found in various non‐nitrogen‐fixing prokaryotes; in particular, they are found in all known methanogens. This indicates that these homologues may play a role in methane production. Here, the cloning of NifH2, a homologue of the NifH nitrogenase component, from Methanocaldococcus jannaschii (MjNifH2) and its expression in Escherichia coli with a polyhistidine tag, purification and crystallization are described. MjNifH2 crystals were obtained by the hanging‐drop vapour‐diffusion method and diffracted to a resolution limit of 2.85 Å. The crystals belonged to space group P2, with unit‐cell parameters a = 64.01, b = 94.38, c = 98.08 Å, α = γ = 90, β = 98.85°. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Acta Crystallographica Section F Wiley

Cloning, expression, purification, crystallization and preliminary crystallographic analysis of NifH2 from Methanocaldococcus jannaschii

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References (2)

Publisher
Wiley
Copyright
International Union of Crystallography, 2011
ISSN
1744-3091
eISSN
1744-3091
DOI
10.1107/S1744309110048104
pmid
21206044
Publisher site
See Article on Publisher Site

Abstract

Nitrogenases are protein complexes that are only found in Azotobacter and are required for biological nitrogen fixation. They are made up of a nitrogenase, which is a NifD2/NifK2 heterotetramer, and a nitrogenase reductase, which is a homodimer of NifH. Many homologues of nitrogenase have been found in various non‐nitrogen‐fixing prokaryotes; in particular, they are found in all known methanogens. This indicates that these homologues may play a role in methane production. Here, the cloning of NifH2, a homologue of the NifH nitrogenase component, from Methanocaldococcus jannaschii (MjNifH2) and its expression in Escherichia coli with a polyhistidine tag, purification and crystallization are described. MjNifH2 crystals were obtained by the hanging‐drop vapour‐diffusion method and diffracted to a resolution limit of 2.85 Å. The crystals belonged to space group P2, with unit‐cell parameters a = 64.01, b = 94.38, c = 98.08 Å, α = γ = 90, β = 98.85°.

Journal

Acta Crystallographica Section FWiley

Published: Jan 1, 2011

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