Proximal ligand tunes active site structure and reactivity in bacterial L. monocytogenes coproheme ferrochelatase

Autores
Dali, Andrea; Sebastiani, Federico; Gabler, Thomas; Frattini, Gianfranco; Moreno, Diego Martin; Estrin, Dario Ariel; Becucci, Maurizio; Hofbauer, Stefan; Smulevich, Giulietta
Año de publicación
2024
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Ferrochelatases catalyze the insertion of ferrous iron into the porphyrin during the heme b biosynthesis pathway, which is fundamental for both prokaryotes and eukaryotes. Interestingly, in the active site of ferrochelatases, the proximal ligand coordinating the porphyrin iron of the product is not conserved, and its catalytic role is still unclear. Here we compare the L. monocytogenes bacterial coproporphyrin ferrochelatase native enzyme together with selected variants, where the proximal Tyr residue was replaced by a His (i.e. the most common ligand in heme proteins), a Met or a Phe (as in human and actinobacterial ferrochelatases, respectively), in their Fe(III), Fe(II) and Fe(II)–CO adduct forms. The study of the active site structure and the activity of the proteins in solution has been performed by UV–vis electronic absorption and resonance Raman spectroscopies, biochemical characterization, and classical MD simulations.All the mutations alter the H-bond interactions between the iron porphyrin propionate groups and the protein, and induce effects on the activity, depending on the polarity of the proximal ligand. The overall results confirm that the weak or non-existing coordination of the porphyrin iron by the proximal residue is essential for the binding of the substrate and the release of the final product.
Fil: Dali, Andrea. Università degli Studi di Firenze; Italia
Fil: Sebastiani, Federico. Università degli Studi di Firenze; Italia
Fil: Gabler, Thomas. University Of Natural Resources And Life Sciences (boku);
Fil: Frattini, Gianfranco. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Química Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Química Rosario; Argentina. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas; Argentina
Fil: Moreno, Diego Martin. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Química Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Química Rosario; Argentina. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas; Argentina
Fil: Estrin, Dario Ariel. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química, Física de los Materiales, Medioambiente y Energía. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química, Física de los Materiales, Medioambiente y Energía; Argentina
Fil: Becucci, Maurizio. Università degli Studi di Firenze; Italia
Fil: Hofbauer, Stefan. University Of Natural Resources And Life Sciences (boku);
Fil: Smulevich, Giulietta. Università degli Studi di Firenze; Italia
Materia
Resonance Raman
Heme biosynthesis
Molecular dynamics simulation
Kinetics
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/267337
Acceder
id CONICETDig_b26bb7d3e5c1354d3927963d60b2d91e
oai_identifier_str oai:ri.conicet.gov.ar:11336/267337
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Proximal ligand tunes active site structure and reactivity in bacterial L. monocytogenes coproheme ferrochelataseDali, AndreaSebastiani, FedericoGabler, ThomasFrattini, GianfrancoMoreno, Diego MartinEstrin, Dario ArielBecucci, MaurizioHofbauer, StefanSmulevich, GiuliettaResonance RamanHeme biosynthesisMolecular dynamics simulationKineticshttps://purl.org/becyt/ford/1.4https://purl.org/becyt/ford/1Ferrochelatases catalyze the insertion of ferrous iron into the porphyrin during the heme b biosynthesis pathway, which is fundamental for both prokaryotes and eukaryotes. Interestingly, in the active site of ferrochelatases, the proximal ligand coordinating the porphyrin iron of the product is not conserved, and its catalytic role is still unclear. Here we compare the L. monocytogenes bacterial coproporphyrin ferrochelatase native enzyme together with selected variants, where the proximal Tyr residue was replaced by a His (i.e. the most common ligand in heme proteins), a Met or a Phe (as in human and actinobacterial ferrochelatases, respectively), in their Fe(III), Fe(II) and Fe(II)–CO adduct forms. The study of the active site structure and the activity of the proteins in solution has been performed by UV–vis electronic absorption and resonance Raman spectroscopies, biochemical characterization, and classical MD simulations.All the mutations alter the H-bond interactions between the iron porphyrin propionate groups and the protein, and induce effects on the activity, depending on the polarity of the proximal ligand. The overall results confirm that the weak or non-existing coordination of the porphyrin iron by the proximal residue is essential for the binding of the substrate and the release of the final product.Fil: Dali, Andrea. Università degli Studi di Firenze; ItaliaFil: Sebastiani, Federico. Università degli Studi di Firenze; ItaliaFil: Gabler, Thomas. University Of Natural Resources And Life Sciences (boku);Fil: Frattini, Gianfranco. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Química Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Química Rosario; Argentina. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas; ArgentinaFil: Moreno, Diego Martin. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Química Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Química Rosario; Argentina. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas; ArgentinaFil: Estrin, Dario Ariel. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química, Física de los Materiales, Medioambiente y Energía. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química, Física de los Materiales, Medioambiente y Energía; ArgentinaFil: Becucci, Maurizio. Università degli Studi di Firenze; ItaliaFil: Hofbauer, Stefan. University Of Natural Resources And Life Sciences (boku);Fil: Smulevich, Giulietta. Università degli Studi di Firenze; ItaliaPergamon-Elsevier Science Ltd2024-05info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/267337Dali, Andrea; Sebastiani, Federico; Gabler, Thomas; Frattini, Gianfranco; Moreno, Diego Martin; et al.; Proximal ligand tunes active site structure and reactivity in bacterial L. monocytogenes coproheme ferrochelatase; Pergamon-Elsevier Science Ltd; Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy; 313; 5-2024; 1-141386-1425CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://linkinghub.elsevier.com/retrieve/pii/S1386142524002865info:eu-repo/semantics/altIdentifier/doi/10.1016/j.saa.2024.124120info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-10-15T14:21:13Zoai:ri.conicet.gov.ar:11336/267337instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-10-15 14:21:14.0CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Proximal ligand tunes active site structure and reactivity in bacterial L. monocytogenes coproheme ferrochelatase
title Proximal ligand tunes active site structure and reactivity in bacterial L. monocytogenes coproheme ferrochelatase
spellingShingle Proximal ligand tunes active site structure and reactivity in bacterial L. monocytogenes coproheme ferrochelatase
Dali, Andrea
Resonance Raman
Heme biosynthesis
Molecular dynamics simulation
Kinetics
title_short Proximal ligand tunes active site structure and reactivity in bacterial L. monocytogenes coproheme ferrochelatase
title_full Proximal ligand tunes active site structure and reactivity in bacterial L. monocytogenes coproheme ferrochelatase
title_fullStr Proximal ligand tunes active site structure and reactivity in bacterial L. monocytogenes coproheme ferrochelatase
title_full_unstemmed Proximal ligand tunes active site structure and reactivity in bacterial L. monocytogenes coproheme ferrochelatase
title_sort Proximal ligand tunes active site structure and reactivity in bacterial L. monocytogenes coproheme ferrochelatase
dc.creator.none.fl_str_mv Dali, Andrea
Sebastiani, Federico
Gabler, Thomas
Frattini, Gianfranco
Moreno, Diego Martin
Estrin, Dario Ariel
Becucci, Maurizio
Hofbauer, Stefan
Smulevich, Giulietta
author Dali, Andrea
author_facet Dali, Andrea
Sebastiani, Federico
Gabler, Thomas
Frattini, Gianfranco
Moreno, Diego Martin
Estrin, Dario Ariel
Becucci, Maurizio
Hofbauer, Stefan
Smulevich, Giulietta
author_role author
author2 Sebastiani, Federico
Gabler, Thomas
Frattini, Gianfranco
Moreno, Diego Martin
Estrin, Dario Ariel
Becucci, Maurizio
Hofbauer, Stefan
Smulevich, Giulietta
author2_role author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv Resonance Raman
Heme biosynthesis
Molecular dynamics simulation
Kinetics
topic Resonance Raman
Heme biosynthesis
Molecular dynamics simulation
Kinetics
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.4
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Ferrochelatases catalyze the insertion of ferrous iron into the porphyrin during the heme b biosynthesis pathway, which is fundamental for both prokaryotes and eukaryotes. Interestingly, in the active site of ferrochelatases, the proximal ligand coordinating the porphyrin iron of the product is not conserved, and its catalytic role is still unclear. Here we compare the L. monocytogenes bacterial coproporphyrin ferrochelatase native enzyme together with selected variants, where the proximal Tyr residue was replaced by a His (i.e. the most common ligand in heme proteins), a Met or a Phe (as in human and actinobacterial ferrochelatases, respectively), in their Fe(III), Fe(II) and Fe(II)–CO adduct forms. The study of the active site structure and the activity of the proteins in solution has been performed by UV–vis electronic absorption and resonance Raman spectroscopies, biochemical characterization, and classical MD simulations.All the mutations alter the H-bond interactions between the iron porphyrin propionate groups and the protein, and induce effects on the activity, depending on the polarity of the proximal ligand. The overall results confirm that the weak or non-existing coordination of the porphyrin iron by the proximal residue is essential for the binding of the substrate and the release of the final product.
Fil: Dali, Andrea. Università degli Studi di Firenze; Italia
Fil: Sebastiani, Federico. Università degli Studi di Firenze; Italia
Fil: Gabler, Thomas. University Of Natural Resources And Life Sciences (boku);
Fil: Frattini, Gianfranco. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Química Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Química Rosario; Argentina. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas; Argentina
Fil: Moreno, Diego Martin. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Química Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Química Rosario; Argentina. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas; Argentina
Fil: Estrin, Dario Ariel. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química, Física de los Materiales, Medioambiente y Energía. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química, Física de los Materiales, Medioambiente y Energía; Argentina
Fil: Becucci, Maurizio. Università degli Studi di Firenze; Italia
Fil: Hofbauer, Stefan. University Of Natural Resources And Life Sciences (boku);
Fil: Smulevich, Giulietta. Università degli Studi di Firenze; Italia
description Ferrochelatases catalyze the insertion of ferrous iron into the porphyrin during the heme b biosynthesis pathway, which is fundamental for both prokaryotes and eukaryotes. Interestingly, in the active site of ferrochelatases, the proximal ligand coordinating the porphyrin iron of the product is not conserved, and its catalytic role is still unclear. Here we compare the L. monocytogenes bacterial coproporphyrin ferrochelatase native enzyme together with selected variants, where the proximal Tyr residue was replaced by a His (i.e. the most common ligand in heme proteins), a Met or a Phe (as in human and actinobacterial ferrochelatases, respectively), in their Fe(III), Fe(II) and Fe(II)–CO adduct forms. The study of the active site structure and the activity of the proteins in solution has been performed by UV–vis electronic absorption and resonance Raman spectroscopies, biochemical characterization, and classical MD simulations.All the mutations alter the H-bond interactions between the iron porphyrin propionate groups and the protein, and induce effects on the activity, depending on the polarity of the proximal ligand. The overall results confirm that the weak or non-existing coordination of the porphyrin iron by the proximal residue is essential for the binding of the substrate and the release of the final product.
publishDate 2024
dc.date.none.fl_str_mv 2024-05
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/267337
Dali, Andrea; Sebastiani, Federico; Gabler, Thomas; Frattini, Gianfranco; Moreno, Diego Martin; et al.; Proximal ligand tunes active site structure and reactivity in bacterial L. monocytogenes coproheme ferrochelatase; Pergamon-Elsevier Science Ltd; Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy; 313; 5-2024; 1-14
1386-1425
CONICET Digital
CONICET
url http://hdl.handle.net/11336/267337
identifier_str_mv Dali, Andrea; Sebastiani, Federico; Gabler, Thomas; Frattini, Gianfranco; Moreno, Diego Martin; et al.; Proximal ligand tunes active site structure and reactivity in bacterial L. monocytogenes coproheme ferrochelatase; Pergamon-Elsevier Science Ltd; Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy; 313; 5-2024; 1-14
1386-1425
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/https://linkinghub.elsevier.com/retrieve/pii/S1386142524002865
info:eu-repo/semantics/altIdentifier/doi/10.1016/j.saa.2024.124120
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
dc.publisher.none.fl_str_mv Pergamon-Elsevier Science Ltd
publisher.none.fl_str_mv Pergamon-Elsevier Science Ltd
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
_version_ 1846082598531497984
score 13.22299

Publicaciones similares