Heme pocket structural properties of a bacterial truncated hemoglobin from thermobifida fusca
- Autores
- Droghetti, Enrica; Nicoletti, Francesco Paolo; Bonamore, Alessandra; Boechi, Leonardo; Arroyo Mañez, Pau; Estrin, Dario Ariel; Boffi, Alberto; Smulevich, Giulietta; Feis, Alessandro
- Año de publicación
- 2010
- Idioma
- inglés
- Tipo de recurso
- artículo
- Estado
- versión publicada
- Descripción
- An acidic surface variant (ASV) of the "truncated" hemoglobin from Thermobifida fusca was designed with the aim of creating a versatile globin scaffold endowed with thermostability and a high level of recombinant expression in its soluble form while keeping the active site unmodified. This engineered protein was obtained by mutating the surface-exposed residues Phe107 and Arg91 to Glu. Molecular dynamics simulations showed that the mutated residues remain solvent-exposed, not affecting the overall protein structure. Thus, the ASV was used in a combinatorial mutagenesis of the distal heme pocket residues in which one, two, or three of the conserved polar residues [TyrB10(54), TyrCD1(67), and TrpG8(119)] were substituted with Phe. Mutants were characterized by infrared and resonance Raman spectroscopy and compared with the wild-type protein. Similar Fe-proximal His stretching frequencies suggest that none of the mutations alters the proximal side of the heme cavity. Two conformers were observed in the spectra of the CO complexes of both wild-type and ASV protein: form 1 with v(FeC) and v(CO) at 509 and 1938 cm-1 and form 2 with v(FeC) and v(CO) at 518 and 1920 cm-1, respectively. Molecular dynamics simulations were performed for the wild-type and ASV forms, as well as for the TyrB10 mutant. The spectroscopic and computational results demonstrate that CO interacts with TrpG8 in form 1 and interacts with both TrpG8 and TyrCD1 in form 2. TyrB10 does not directly interact with the bound CO.
Fil: Droghetti, Enrica. Universita di Firenze; Italia
Fil: Nicoletti, Francesco Paolo. Universita di Firenze; Italia
Fil: Bonamore, Alessandra. Università degli studi di Roma "La Sapienza"; Italia
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
Fil: Arroyo Mañez, Pau. 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
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
Fil: Boffi, Alberto. Università degli studi di Roma "La Sapienza"; Italia
Fil: Smulevich, Giulietta. Universita di Firenze; Italia
Fil: Feis, Alessandro. Universita di Firenze; Italia - Materia
-
Structural Properties
Truncated Hemoglobin
Thermofida Fusca - Nivel de accesibilidad
- acceso abierto
- Condiciones de uso
- https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
- Repositorio
- Institución
- Consejo Nacional de Investigaciones Científicas y Técnicas
- OAI Identificador
- oai:ri.conicet.gov.ar:11336/83871
Ver los metadatos del registro completo
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Heme pocket structural properties of a bacterial truncated hemoglobin from thermobifida fuscaDroghetti, EnricaNicoletti, Francesco PaoloBonamore, AlessandraBoechi, LeonardoArroyo Mañez, PauEstrin, Dario ArielBoffi, AlbertoSmulevich, GiuliettaFeis, AlessandroStructural PropertiesTruncated HemoglobinThermofida Fuscahttps://purl.org/becyt/ford/1.4https://purl.org/becyt/ford/1An acidic surface variant (ASV) of the "truncated" hemoglobin from Thermobifida fusca was designed with the aim of creating a versatile globin scaffold endowed with thermostability and a high level of recombinant expression in its soluble form while keeping the active site unmodified. This engineered protein was obtained by mutating the surface-exposed residues Phe107 and Arg91 to Glu. Molecular dynamics simulations showed that the mutated residues remain solvent-exposed, not affecting the overall protein structure. Thus, the ASV was used in a combinatorial mutagenesis of the distal heme pocket residues in which one, two, or three of the conserved polar residues [TyrB10(54), TyrCD1(67), and TrpG8(119)] were substituted with Phe. Mutants were characterized by infrared and resonance Raman spectroscopy and compared with the wild-type protein. Similar Fe-proximal His stretching frequencies suggest that none of the mutations alters the proximal side of the heme cavity. Two conformers were observed in the spectra of the CO complexes of both wild-type and ASV protein: form 1 with v(FeC) and v(CO) at 509 and 1938 cm-1 and form 2 with v(FeC) and v(CO) at 518 and 1920 cm-1, respectively. Molecular dynamics simulations were performed for the wild-type and ASV forms, as well as for the TyrB10 mutant. The spectroscopic and computational results demonstrate that CO interacts with TrpG8 in form 1 and interacts with both TrpG8 and TyrCD1 in form 2. TyrB10 does not directly interact with the bound CO.Fil: Droghetti, Enrica. Universita di Firenze; ItaliaFil: Nicoletti, Francesco Paolo. Universita di Firenze; ItaliaFil: Bonamore, Alessandra. Università degli studi di Roma "La Sapienza"; ItaliaFil: 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; ArgentinaFil: Arroyo Mañez, Pau. 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; 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; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Inorgánica, Analítica y Química Física; ArgentinaFil: Boffi, Alberto. Università degli studi di Roma "La Sapienza"; ItaliaFil: Smulevich, Giulietta. Universita di Firenze; ItaliaFil: Feis, Alessandro. Universita di Firenze; ItaliaAmerican Chemical Society2010-12info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/83871Droghetti, Enrica; Nicoletti, Francesco Paolo; Bonamore, Alessandra; Boechi, Leonardo; Arroyo Mañez, Pau; et al.; Heme pocket structural properties of a bacterial truncated hemoglobin from thermobifida fusca; American Chemical Society; Biochemistry; 49; 49; 12-2010; 10394-104020006-2960CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.1021/bi101452kinfo:eu-repo/semantics/altIdentifier/url/https://pubs.acs.org/doi/10.1021/bi101452kinfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T09:43:26Zoai:ri.conicet.gov.ar:11336/83871instacron: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:34982025-09-29 09:43:26.72CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse |
dc.title.none.fl_str_mv |
Heme pocket structural properties of a bacterial truncated hemoglobin from thermobifida fusca |
title |
Heme pocket structural properties of a bacterial truncated hemoglobin from thermobifida fusca |
spellingShingle |
Heme pocket structural properties of a bacterial truncated hemoglobin from thermobifida fusca Droghetti, Enrica Structural Properties Truncated Hemoglobin Thermofida Fusca |
title_short |
Heme pocket structural properties of a bacterial truncated hemoglobin from thermobifida fusca |
title_full |
Heme pocket structural properties of a bacterial truncated hemoglobin from thermobifida fusca |
title_fullStr |
Heme pocket structural properties of a bacterial truncated hemoglobin from thermobifida fusca |
title_full_unstemmed |
Heme pocket structural properties of a bacterial truncated hemoglobin from thermobifida fusca |
title_sort |
Heme pocket structural properties of a bacterial truncated hemoglobin from thermobifida fusca |
dc.creator.none.fl_str_mv |
Droghetti, Enrica Nicoletti, Francesco Paolo Bonamore, Alessandra Boechi, Leonardo Arroyo Mañez, Pau Estrin, Dario Ariel Boffi, Alberto Smulevich, Giulietta Feis, Alessandro |
author |
Droghetti, Enrica |
author_facet |
Droghetti, Enrica Nicoletti, Francesco Paolo Bonamore, Alessandra Boechi, Leonardo Arroyo Mañez, Pau Estrin, Dario Ariel Boffi, Alberto Smulevich, Giulietta Feis, Alessandro |
author_role |
author |
author2 |
Nicoletti, Francesco Paolo Bonamore, Alessandra Boechi, Leonardo Arroyo Mañez, Pau Estrin, Dario Ariel Boffi, Alberto Smulevich, Giulietta Feis, Alessandro |
author2_role |
author author author author author author author author |
dc.subject.none.fl_str_mv |
Structural Properties Truncated Hemoglobin Thermofida Fusca |
topic |
Structural Properties Truncated Hemoglobin Thermofida Fusca |
purl_subject.fl_str_mv |
https://purl.org/becyt/ford/1.4 https://purl.org/becyt/ford/1 |
dc.description.none.fl_txt_mv |
An acidic surface variant (ASV) of the "truncated" hemoglobin from Thermobifida fusca was designed with the aim of creating a versatile globin scaffold endowed with thermostability and a high level of recombinant expression in its soluble form while keeping the active site unmodified. This engineered protein was obtained by mutating the surface-exposed residues Phe107 and Arg91 to Glu. Molecular dynamics simulations showed that the mutated residues remain solvent-exposed, not affecting the overall protein structure. Thus, the ASV was used in a combinatorial mutagenesis of the distal heme pocket residues in which one, two, or three of the conserved polar residues [TyrB10(54), TyrCD1(67), and TrpG8(119)] were substituted with Phe. Mutants were characterized by infrared and resonance Raman spectroscopy and compared with the wild-type protein. Similar Fe-proximal His stretching frequencies suggest that none of the mutations alters the proximal side of the heme cavity. Two conformers were observed in the spectra of the CO complexes of both wild-type and ASV protein: form 1 with v(FeC) and v(CO) at 509 and 1938 cm-1 and form 2 with v(FeC) and v(CO) at 518 and 1920 cm-1, respectively. Molecular dynamics simulations were performed for the wild-type and ASV forms, as well as for the TyrB10 mutant. The spectroscopic and computational results demonstrate that CO interacts with TrpG8 in form 1 and interacts with both TrpG8 and TyrCD1 in form 2. TyrB10 does not directly interact with the bound CO. Fil: Droghetti, Enrica. Universita di Firenze; Italia Fil: Nicoletti, Francesco Paolo. Universita di Firenze; Italia Fil: Bonamore, Alessandra. Università degli studi di Roma "La Sapienza"; Italia 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 Fil: Arroyo Mañez, Pau. 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 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 Fil: Boffi, Alberto. Università degli studi di Roma "La Sapienza"; Italia Fil: Smulevich, Giulietta. Universita di Firenze; Italia Fil: Feis, Alessandro. Universita di Firenze; Italia |
description |
An acidic surface variant (ASV) of the "truncated" hemoglobin from Thermobifida fusca was designed with the aim of creating a versatile globin scaffold endowed with thermostability and a high level of recombinant expression in its soluble form while keeping the active site unmodified. This engineered protein was obtained by mutating the surface-exposed residues Phe107 and Arg91 to Glu. Molecular dynamics simulations showed that the mutated residues remain solvent-exposed, not affecting the overall protein structure. Thus, the ASV was used in a combinatorial mutagenesis of the distal heme pocket residues in which one, two, or three of the conserved polar residues [TyrB10(54), TyrCD1(67), and TrpG8(119)] were substituted with Phe. Mutants were characterized by infrared and resonance Raman spectroscopy and compared with the wild-type protein. Similar Fe-proximal His stretching frequencies suggest that none of the mutations alters the proximal side of the heme cavity. Two conformers were observed in the spectra of the CO complexes of both wild-type and ASV protein: form 1 with v(FeC) and v(CO) at 509 and 1938 cm-1 and form 2 with v(FeC) and v(CO) at 518 and 1920 cm-1, respectively. Molecular dynamics simulations were performed for the wild-type and ASV forms, as well as for the TyrB10 mutant. The spectroscopic and computational results demonstrate that CO interacts with TrpG8 in form 1 and interacts with both TrpG8 and TyrCD1 in form 2. TyrB10 does not directly interact with the bound CO. |
publishDate |
2010 |
dc.date.none.fl_str_mv |
2010-12 |
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/11336/83871 Droghetti, Enrica; Nicoletti, Francesco Paolo; Bonamore, Alessandra; Boechi, Leonardo; Arroyo Mañez, Pau; et al.; Heme pocket structural properties of a bacterial truncated hemoglobin from thermobifida fusca; American Chemical Society; Biochemistry; 49; 49; 12-2010; 10394-10402 0006-2960 CONICET Digital CONICET |
url |
http://hdl.handle.net/11336/83871 |
identifier_str_mv |
Droghetti, Enrica; Nicoletti, Francesco Paolo; Bonamore, Alessandra; Boechi, Leonardo; Arroyo Mañez, Pau; et al.; Heme pocket structural properties of a bacterial truncated hemoglobin from thermobifida fusca; American Chemical Society; Biochemistry; 49; 49; 12-2010; 10394-10402 0006-2960 CONICET Digital CONICET |
dc.language.none.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
info:eu-repo/semantics/altIdentifier/doi/10.1021/bi101452k info:eu-repo/semantics/altIdentifier/url/https://pubs.acs.org/doi/10.1021/bi101452k |
dc.rights.none.fl_str_mv |
info:eu-repo/semantics/openAccess https://creativecommons.org/licenses/by-nc-sa/2.5/ar/ |
eu_rights_str_mv |
openAccess |
rights_invalid_str_mv |
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/ |
dc.format.none.fl_str_mv |
application/pdf application/pdf application/pdf application/pdf |
dc.publisher.none.fl_str_mv |
American Chemical Society |
publisher.none.fl_str_mv |
American Chemical Society |
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) |
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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 |
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score |
13.070432 |