Unique Biradical Intermediate in the Mechanism of the Heme Enzyme Chlorite Dismutase
| Autor(a) principal: | |
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| Data de Publicação: | 2021 |
| Outros Autores: | , , , , , , |
| Tipo de documento: | Artigo |
| Idioma: | eng |
| Título da fonte: | Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
| Texto Completo: | http://hdl.handle.net/10362/142216 |
Resumo: | Funding 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. |
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Unique Biradical Intermediate in the Mechanism of the Heme Enzyme Chlorite Dismutasebiradicalchlorite dismutaseelectron paramagnetic resonance spectroscopyhememicrosecond-timescale freeze hyperquenchingrapid kineticsresonance Raman spectroscopytriplet stateCatalysisChemistry(all)Funding 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.The 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.Instituto de Tecnologia Química e Biológica António Xavier (ITQB)RUNPüschmann, JuliaMahor, DurgaDe Geus, Daniël C.Strampraad, Marc J.F.Srour, BatoulHagen, Wilfred R.Todorovic, SmiljaHagedoorn, Peter Leon2022-07-20T22:30:19Z2021-12-032021-12-03T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article12application/pdfhttp://hdl.handle.net/10362/142216eng2155-5435PURE: 42409043https://doi.org/10.1021/acscatal.1c03432info:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2024-03-11T05:19:43Zoai:run.unl.pt:10362/142216Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-20T03:50:12.253391Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse |
| dc.title.none.fl_str_mv |
Unique Biradical Intermediate in the Mechanism of the Heme Enzyme Chlorite Dismutase |
| title |
Unique Biradical Intermediate in the Mechanism of the Heme Enzyme Chlorite Dismutase |
| spellingShingle |
Unique Biradical Intermediate in the Mechanism of the Heme Enzyme Chlorite Dismutase Püschmann, Julia biradical chlorite dismutase electron paramagnetic resonance spectroscopy heme microsecond-timescale freeze hyperquenching rapid kinetics resonance Raman spectroscopy triplet state Catalysis Chemistry(all) |
| title_short |
Unique Biradical Intermediate in the Mechanism of the Heme Enzyme Chlorite Dismutase |
| title_full |
Unique Biradical Intermediate in the Mechanism of the Heme Enzyme Chlorite Dismutase |
| title_fullStr |
Unique Biradical Intermediate in the Mechanism of the Heme Enzyme Chlorite Dismutase |
| title_full_unstemmed |
Unique Biradical Intermediate in the Mechanism of the Heme Enzyme Chlorite Dismutase |
| title_sort |
Unique Biradical Intermediate in the Mechanism of the Heme Enzyme Chlorite Dismutase |
| author |
Püschmann, Julia |
| author_facet |
Püschmann, Julia Mahor, Durga De Geus, Daniël C. Strampraad, Marc J.F. Srour, Batoul Hagen, Wilfred R. Todorovic, Smilja Hagedoorn, Peter Leon |
| author_role |
author |
| author2 |
Mahor, Durga De Geus, Daniël C. Strampraad, Marc J.F. Srour, Batoul Hagen, Wilfred R. Todorovic, Smilja Hagedoorn, Peter Leon |
| author2_role |
author author author author author author author |
| dc.contributor.none.fl_str_mv |
Instituto de Tecnologia Química e Biológica António Xavier (ITQB) RUN |
| dc.contributor.author.fl_str_mv |
Püschmann, Julia Mahor, Durga De Geus, Daniël C. Strampraad, Marc J.F. Srour, Batoul Hagen, Wilfred R. Todorovic, Smilja Hagedoorn, Peter Leon |
| dc.subject.por.fl_str_mv |
biradical chlorite dismutase electron paramagnetic resonance spectroscopy heme microsecond-timescale freeze hyperquenching rapid kinetics resonance Raman spectroscopy triplet state Catalysis Chemistry(all) |
| topic |
biradical chlorite dismutase electron paramagnetic resonance spectroscopy heme microsecond-timescale freeze hyperquenching rapid kinetics resonance Raman spectroscopy triplet state Catalysis Chemistry(all) |
| description |
Funding 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. |
| publishDate |
2021 |
| dc.date.none.fl_str_mv |
2021-12-03 2021-12-03T00:00:00Z 2022-07-20T22:30:19Z |
| dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
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info:eu-repo/semantics/article |
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article |
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publishedVersion |
| dc.identifier.uri.fl_str_mv |
http://hdl.handle.net/10362/142216 |
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http://hdl.handle.net/10362/142216 |
| dc.language.iso.fl_str_mv |
eng |
| language |
eng |
| dc.relation.none.fl_str_mv |
2155-5435 PURE: 42409043 https://doi.org/10.1021/acscatal.1c03432 |
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info:eu-repo/semantics/openAccess |
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openAccess |
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12 application/pdf |
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Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
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RCAAP |
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RCAAP |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
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