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dc.contributor.author
Pesce, Alessandra
dc.contributor.author
Bustamante, Juan Pablo
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Bidon Chanal, Axel
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Boechi, Leonardo
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Estrin, Dario Ariel
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Luque, Francisco Javier
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Sebilo, Anne
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Guertin, Michel
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Bolognesi, Martino
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Ascenzi, Paolo
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Nardini, Marco
dc.date.available
2018-09-10T19:34:41Z
dc.date.issued
2016-01
dc.identifier.citation
Pesce, Alessandra; Bustamante, Juan Pablo; Bidon Chanal, Axel; Boechi, Leonardo; Estrin, Dario Ariel; et al.; The N-terminal pre-A region of Mycobacterium tuberculosis 2/2HbN promotes NO-dioxygenase activity; Wiley Blackwell Publishing, Inc; Febs Journal; 283; 2; 1-2016; 305-322
dc.identifier.issn
1742-464X
dc.identifier.uri
http://hdl.handle.net/11336/58965
dc.description.abstract
A unique defense mechanisms by which Mycobacterium tuberculosis protects itself from nitrosative stress is based on the O2-dependent NO-dioxygenase (NOD) activity of truncated hemoglobin 2/2HbN (Mt2/2HbN). The NOD activity largely depends on the efficiency of ligand migration to the heme cavity through a two-tunnel (long and short) system; recently, it was also correlated with the presence at the Mt2/2HbN N-terminus of a short pre-A region, not conserved in most 2/2HbNs, whose deletion results in a drastic reduction of NO scavenging. In the present study, we report the crystal structure of Mt2/2HbN-ΔpreA, lacking the pre-A region, at a resolution of 1.53 Å. We show that removal of the pre-A region results in long range effects on the protein C-terminus, promoting the assembly of a stable dimer, both in the crystals and in solution. In the Mt2/2HbN-ΔpreA dimer, access of heme ligands to the short tunnel is hindered. Molecular dynamics simulations show that the long tunnel branch is the only accessible pathway for O2-ligand migration to/from the heme, and that the gating residue Phe(62)E15 partly restricts the diameter of the tunnel. Accordingly, kinetic measurements indicate that the kon value for peroxynitrite isomerization by Mt2/2HbN-ΔpreA-Fe(III) is four-fold lower relative to the full-length protein, and that NO scavenging by Mt2/2HbN-ΔpreA-Fe(II)-O2 is reduced by 35-fold. Therefore, we speculate that Mt2/2HbN evolved to host the pre-A region as a mechanism for preventing dimerization, thus reinforcing the survival of the microorganism against the reactive nitrosative stress in macrophages. Database Coordinates and structure factors have been deposited in the Protein Data Bank under accession number 5AB8. Removal of the pre-A region in M. tuberculosis 2/2HbN (Mt2/2HbN-ΔpreA) results in dimerization and in a reduced access to the heme pocket. Kinetic measurements indicate a 4-fold decrease in kon for peroxynitrite isomerization and a 35-fold decrease in NO-scavenging relative to full-length Mt2/2HbN. Thus, the pre-A region might be involved in reinforcing survival of the microorganism against nitrosative stress in macrophages.
dc.format
application/pdf
dc.language.iso
eng
dc.publisher
Wiley Blackwell Publishing, Inc
dc.rights
info:eu-repo/semantics/openAccess
dc.rights.uri
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.subject
2/2 Hemoglobins
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Globin Dynamics
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Heme/Ligand Tunneling
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No Dioxygenase
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Truncated Hemoglobins
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Otras Ciencias Químicas
dc.subject.classification
Ciencias Químicas
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CIENCIAS NATURALES Y EXACTAS
dc.title
The N-terminal pre-A region of Mycobacterium tuberculosis 2/2HbN promotes NO-dioxygenase activity
dc.type
info:eu-repo/semantics/article
dc.type
info:ar-repo/semantics/artículo
dc.type
info:eu-repo/semantics/publishedVersion
dc.date.updated
2018-09-04T19:05:23Z
dc.journal.volume
283
dc.journal.number
2
dc.journal.pagination
305-322
dc.journal.pais
Reino Unido
dc.journal.ciudad
Londres
dc.description.fil
Fil: Pesce, Alessandra. Università degli Studi di Genova; Italia
dc.description.fil
Fil: Bustamante, Juan Pablo. 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. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Inorgánica, Analítica y Química Física; Argentina
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Fil: Bidon Chanal, Axel. Universidad de Barcelona; España
dc.description.fil
Fil: Boechi, Leonardo. 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. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Inorgánica, Analítica y Química Física; Argentina
dc.description.fil
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. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Inorgánica, Analítica y Química Física; Argentina
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Fil: Luque, Francisco Javier. Universidad de Barcelona; España
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Fil: Sebilo, Anne. Laval University; Canadá
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Fil: Guertin, Michel. Laval University; Canadá
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Fil: Bolognesi, Martino. Universidad de Milan; Italia
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Fil: Ascenzi, Paolo. Università di Roma; Italia
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Fil: Nardini, Marco. Universidad de Milan; Italia
dc.journal.title
Febs Journal
dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/doi/https://dx.doi.org/10.1111/febs.13571
dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/url/https://febs.onlinelibrary.wiley.com/doi/abs/10.1111/febs.13571
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