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Unique Biradical Intermediate in the Mechanism of the Heme Enzyme Chlorite Dismutase

2021-12-03Artigo científico Acesso aberto
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dc.contributor.authorPüschmann, Julia
dc.contributor.authorMahor, Durga
dc.contributor.authorDe Geus, Daniël C.
dc.contributor.authorStrampraad, Marc J.F.
dc.contributor.authorSrour, Batoul
dc.contributor.authorHagen, Wilfred R.
dc.contributor.authorTodorovic, Smilja
dc.contributor.authorHagedoorn, Peter Leon
dc.contributor.institutionInstituto de Tecnologia Química e Biológica António Xavier (ITQB)
dc.contributor.pblACS - American Chemical Society
dc.date.accessioned2022-07-20T22:30:19Z
dc.date.available2022-07-20T22:30:19Z
dc.date.issued2021-12-03
dc.descriptionFunding Information: This research was supported by the grant NWO–CW 711.014.006 from the Council for Chemical Sciences of The Netherlands Organization for Scientific Research. S.T. acknowledges the support from the Project LISBOA-01-0145-FEDER-007660 (Microbiologia Molecular, Estrutural e Celular) funded by FEDER funds through COMPETE 2020─Programa Operacional Competitividade e Internacionalização (POCI) and by national funds through FCT─Fundação para a Ciência e a Tecnologia and from the European Union’s Horizon 2020 Research and Innovation Program through TIMB3 (grant agreements no 810856). This work is dedicated to the late Prof. Simon de Vries, who pioneered the development of the rapid kinetic techniques that have been seminal in this study. Publisher Copyright: © 2021 The Authors. Published by American Chemical Society.
dc.description.abstractThe heme enzyme chlorite dismutase (Cld) catalyzes O-O bond formation as part of the conversion of the toxic chlorite (ClO2-) to chloride (Cl-) and molecular oxygen (O2). Enzymatic O-O bond formation is rare in nature, and therefore, the reaction mechanism of Cld is of great interest. Microsecond timescale pre-steady-state kinetic experiments employing Cld from Azospira oryzae (AoCld), the natural substrate chlorite, and the model substrate peracetic acid (PAA) reveal the formation of distinct intermediates. AoCld forms a complex with PAA rapidly, which is cleaved heterolytically to yield Compound I, which is sequentially converted to Compound II. In the presence of chlorite, AoCld forms an initial intermediate with spectroscopic characteristics of a 6-coordinate high-spin ferric substrate adduct, which subsequently transforms at kobs = 2-5 × 104 s-1 to an intermediate 5-coordinated high-spin ferric species. Microsecond-timescale freeze-hyperquench experiments uncovered the presence of a transient low-spin ferric species and a triplet species attributed to two weakly coupled amino acid cation radicals. The intermediates of the chlorite reaction were not observed with the model substrate PAA. These findings demonstrate the nature of physiologically relevant catalytic intermediates and show that the commonly used model substrate may not behave as expected, which demands a revision of the currently proposed mechanism of Clds. The transient triplet-state biradical species that we designate as Compound T is, to the best of our knowledge, unique in heme enzymology. The results highlight electron paramagnetic resonance spectroscopic evidence for transient intermediate formation during the reaction of AoCld with its natural substrate chlorite. In the proposed mechanism, the heme iron remains ferric throughout the catalytic cycle, which may minimize the heme moiety's reorganization and thereby maximize the enzyme's catalytic efficiency.en
dc.description.versionpublishersversion
dc.description.versionpublished
dc.format.extent12
dc.format.extent2840262
dc.identifier.doi10.1021/acscatal.1c03432
dc.identifier.issn2155-5435
dc.identifier.otherPURE: 42409043
dc.identifier.otherPURE UUID: 28012180-8d50-4998-a131-45412347c651
dc.identifier.otherScopus: 85119976292
dc.identifier.urihttp://hdl.handle.net/10362/142216
dc.identifier.urlhttps://www.scopus.com/pages/publications/85119976292
dc.identifier.urlhttps://pubs.acs.org/doi/pdf/10.1021/acscatal.1c03432
dc.language.isoeng
dc.peerreviewedyes
dc.subjectbiradical
dc.subjectchlorite dismutase
dc.subjectelectron paramagnetic resonance spectroscopy
dc.subjectheme
dc.subjectmicrosecond-timescale freeze hyperquenching
dc.subjectrapid kinetics
dc.subjectresonance Raman spectroscopy
dc.subjecttriplet state
dc.subjectCatalysis
dc.subjectGeneral Chemistry
dc.titleUnique Biradical Intermediate in the Mechanism of the Heme Enzyme Chlorite Dismutaseen
dc.typejournal article
degois.publication.firstPage14533
degois.publication.issue23
degois.publication.lastPage14544
degois.publication.titleACS Catalysis
degois.publication.volume11
dspace.entity.typePublication
rcaap.rightsopenAccess

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