Subforms and In Vitro Reconstitution of the NAD-Reducing Hydrogenase of Alcaligenes eutrophus
10.1128/jb.180.5.1023-1029.1998
Crossref journal-article
American Society for Microbiology
Journal of Bacteriology (235)
Abstract

ABSTRACT The cytoplasmic, NAD-reducing hydrogenase (SH) of Alcaligenes eutrophus H16 is a heterotetrameric enzyme which contains several cofactors and undergoes a complex maturation during biogenesis. HoxH is the Ni-carrying subunit, and together with HoxY it forms the hydrogenase dimer. HoxF and HoxU represent the flavin-containing diaphorase moiety, which is closely related to NADH:ubiquinone oxidoreductase and mediates NADH oxidation. A variety of mutations were introduced into the four SH structural genes to obtain mutant enzymes composed of monomeric and dimeric forms. A deletion removing most of hoxF , hoxU , and hoxY led to the expression of a HoxH monomer derivative which was proteolytically processed at the C terminus like the wild-type polypeptide. While the hydrogenase dimer, produced by a strain deleted of hoxF and hoxU , displayed H 2 -dependent dye-reducing activity, the monomeric form did not mediate the activation of H 2 , although nickel was incorporated into HoxH. Deletion of hoxH and hoxY led to the production of HoxFU dimers which displayed NADH:oxidoreductase activity. Mixing the hydrogenase and the diaphorase moieties in vitro reconstituted the structure and catalytic function of the SH holoenzyme.

Bibliography

Massanz, C., Schmidt, S., & Friedrich, B. (1998). Subforms and In Vitro Reconstitution of the NAD-Reducing Hydrogenase of Alcaligenes eutrophus. Journal of Bacteriology, 180(5), 1023–1029.

Authors 3
  1. Christian Massanz (first)
  2. Silke Schmidt (additional)
  3. Bärbel Friedrich (additional)
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Dates
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Created 5 years, 7 months ago (Dec. 31, 2019, 11:30 a.m.)
Deposited 4 years ago (July 29, 2021, 2:26 p.m.)
Indexed 2 months ago (June 27, 2025, 6:39 a.m.)
Issued 27 years, 5 months ago (March 1, 1998)
Published 27 years, 5 months ago (March 1, 1998)
Published Print 27 years, 5 months ago (March 1, 1998)
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@article{Massanz_1998, title={Subforms and In Vitro Reconstitution of the NAD-Reducing Hydrogenase of Alcaligenes eutrophus}, volume={180}, ISSN={1098-5530}, url={http://dx.doi.org/10.1128/jb.180.5.1023-1029.1998}, DOI={10.1128/jb.180.5.1023-1029.1998}, number={5}, journal={Journal of Bacteriology}, publisher={American Society for Microbiology}, author={Massanz, Christian and Schmidt, Silke and Friedrich, Bärbel}, year={1998}, month=mar, pages={1023–1029} }