A [3Cu:2S] cluster provides insight into the assembly and function of the CuZ site of nitrous oxide reductase

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Abstract: Nitrous oxide reductase (N2OR) is the only known enzyme reducing environmentally critical nitrous oxide (N2O) to dinitrogen (N2) as the final step of bacterial denitrification. The assembly process of its unique catalytic [4Cu:2S] cluster CuZ remains scarcely understood. Here we report on a mutagenesis study of all seven histidine ligands coordinating this copper center, followed by spectroscopic and structural characterization and based on an established, functional expression system for Pseudomonas stutzeri N2OR in Escherichia coli. While no copper ion was found in the CuZ binding site of variants H129A, H130A, H178A, H326A, H433A and H494A, the H382A variant carried a catalytically inactive [3Cu:2S] center, in which one sulfur ligand, SZ2, had relocated to form a weak hydrogen bond to the sidechain of the nearby lysine residue K454. This link provides sufficient stability to avoid the loss of the sulfide anion. The UV-vis spectra of this cluster are strikingly similar to those of the active enzyme, implying that the flexibility of SZ2 may have been observed before, but not recognized. The sulfide shift changes the metal coordination in CuZ and is thus of high mechanistic interest

Location
Deutsche Nationalbibliothek Frankfurt am Main
Extent
Online-Ressource
Language
Englisch
Notes
Chemical science. - 12, 9 (2021) , 3239-3244, ISSN: 2041-6539

Event
Veröffentlichung
(where)
Freiburg
(who)
Universität
(when)
2023
Creator
Zhang, Lin
Bill, Eckhard
Kroneck, Peter M. H.
Einsle, Oliver
Contributor

DOI
10.1039/d0sc05204c
URN
urn:nbn:de:bsz:25-freidok-2364088
Rights
Open Access; Der Zugriff auf das Objekt ist unbeschränkt möglich.
Last update
25.03.2025, 1:54 PM CET

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Time of origin

  • 2023

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