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Determination of the active form of the tetranuclear copper sulfur cluster in nitrous oxide reductase
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Orientador(es)
Resumo(s)
N2OR has been found to have two structural forms of its tetranuclear copper active site, the 4CuS Cu-Z* form and the 4Cu2S Cu-Z form. EPR, resonance Raman, and MCD spectroscopies have been used to determine the redox states of these sites under different reductant conditions, showing that the Cu-Z* site accesses the 1-hole and fully reduced redox states, while the Cu-Z site accesses the 2-hole and 1-hole redox states. Single-turnover reactions of N2OR for Cu-Z and Cu-Z* poised in these redox states and steady-state turnover assays with different proportions of Cu-Z and Cu-Z* show that only fully reduced Cu-Z* is catalytically competent in rapid turnover with N2O.
Descrição
This research was supported by NIH grant DK-31450 (E.I.S.) and National funds by FCT (Fundacao para a Ciencia e a Tecnologia) under the projects PEst-C/EQB/LA0006/2013 and PTDC/QUI-BIQ/116481/2010 (I.M.).
Palavras-chave
CU-Z CLUSTER ACHROMOBACTER-CYCLOCLASTES PSEUDOMONAS-NAUTICA CATALYTIC CENTER N2O REDUCTION PURIFICATION ENZYME RESONANCE MECHANISM INSIGHT