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Nitrogen Fixation and Hydrogen Evolution by Sterically Encumbered Mo-Nitrogenase
[Image: see text] The substrate-reducing proteins of all nitrogenases (MoFe, VFe, and FeFe) are organized as α(2)ß(2)(γ(2)) multimers with two functional halves. While their dimeric organization could afford improved structural stability of nitrogenases in vivo, previous research has proposed both n...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Chemical Society
2023
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10207099/ https://www.ncbi.nlm.nih.gov/pubmed/37234119 http://dx.doi.org/10.1021/jacsau.3c00165 |
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author | Cadoux, Cécile Ratcliff, Daniel Maslać, Nevena Gu, Wenyu Tsakoumagkos, Ioannis Hoogendoorn, Sascha Wagner, Tristan Milton, Ross D. |
author_facet | Cadoux, Cécile Ratcliff, Daniel Maslać, Nevena Gu, Wenyu Tsakoumagkos, Ioannis Hoogendoorn, Sascha Wagner, Tristan Milton, Ross D. |
author_sort | Cadoux, Cécile |
collection | PubMed |
description | [Image: see text] The substrate-reducing proteins of all nitrogenases (MoFe, VFe, and FeFe) are organized as α(2)ß(2)(γ(2)) multimers with two functional halves. While their dimeric organization could afford improved structural stability of nitrogenases in vivo, previous research has proposed both negative and positive cooperativity contributions with respect to enzymatic activity. Here, a 1.4 kDa peptide was covalently introduced in the proximity of the P cluster, corresponding to the Fe protein docking position. The Strep-tag carried by the added peptide simultaneously sterically inhibits electron delivery to the MoFe protein and allows the isolation of partially inhibited MoFe proteins (where the half-inhibited MoFe protein was targeted). We confirm that the partially functional MoFe protein retains its ability to reduce N(2) to NH(3), with no significant difference in selectivity over obligatory/parasitic H(2) formation. Our experiment concludes that wild-type nitrogenase exhibits negative cooperativity during the steady state regarding H(2) and NH(3) formation (under Ar or N(2)), with one-half of the MoFe protein inhibiting turnover in the second half. This emphasizes the presence and importance of long-range (>95 Å) protein–protein communication in biological N(2) fixation in Azotobacter vinelandii. |
format | Online Article Text |
id | pubmed-10207099 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | American Chemical Society |
record_format | MEDLINE/PubMed |
spelling | pubmed-102070992023-05-25 Nitrogen Fixation and Hydrogen Evolution by Sterically Encumbered Mo-Nitrogenase Cadoux, Cécile Ratcliff, Daniel Maslać, Nevena Gu, Wenyu Tsakoumagkos, Ioannis Hoogendoorn, Sascha Wagner, Tristan Milton, Ross D. JACS Au [Image: see text] The substrate-reducing proteins of all nitrogenases (MoFe, VFe, and FeFe) are organized as α(2)ß(2)(γ(2)) multimers with two functional halves. While their dimeric organization could afford improved structural stability of nitrogenases in vivo, previous research has proposed both negative and positive cooperativity contributions with respect to enzymatic activity. Here, a 1.4 kDa peptide was covalently introduced in the proximity of the P cluster, corresponding to the Fe protein docking position. The Strep-tag carried by the added peptide simultaneously sterically inhibits electron delivery to the MoFe protein and allows the isolation of partially inhibited MoFe proteins (where the half-inhibited MoFe protein was targeted). We confirm that the partially functional MoFe protein retains its ability to reduce N(2) to NH(3), with no significant difference in selectivity over obligatory/parasitic H(2) formation. Our experiment concludes that wild-type nitrogenase exhibits negative cooperativity during the steady state regarding H(2) and NH(3) formation (under Ar or N(2)), with one-half of the MoFe protein inhibiting turnover in the second half. This emphasizes the presence and importance of long-range (>95 Å) protein–protein communication in biological N(2) fixation in Azotobacter vinelandii. American Chemical Society 2023-05-09 /pmc/articles/PMC10207099/ /pubmed/37234119 http://dx.doi.org/10.1021/jacsau.3c00165 Text en © 2023 The Authors. Published by American Chemical Society https://creativecommons.org/licenses/by/4.0/Permits the broadest form of re-use including for commercial purposes, provided that author attribution and integrity are maintained (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Cadoux, Cécile Ratcliff, Daniel Maslać, Nevena Gu, Wenyu Tsakoumagkos, Ioannis Hoogendoorn, Sascha Wagner, Tristan Milton, Ross D. Nitrogen Fixation and Hydrogen Evolution by Sterically Encumbered Mo-Nitrogenase |
title | Nitrogen Fixation and Hydrogen Evolution by Sterically
Encumbered Mo-Nitrogenase |
title_full | Nitrogen Fixation and Hydrogen Evolution by Sterically
Encumbered Mo-Nitrogenase |
title_fullStr | Nitrogen Fixation and Hydrogen Evolution by Sterically
Encumbered Mo-Nitrogenase |
title_full_unstemmed | Nitrogen Fixation and Hydrogen Evolution by Sterically
Encumbered Mo-Nitrogenase |
title_short | Nitrogen Fixation and Hydrogen Evolution by Sterically
Encumbered Mo-Nitrogenase |
title_sort | nitrogen fixation and hydrogen evolution by sterically
encumbered mo-nitrogenase |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10207099/ https://www.ncbi.nlm.nih.gov/pubmed/37234119 http://dx.doi.org/10.1021/jacsau.3c00165 |
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