A spectroscopy-based methodology for rapid screening and characterization of phytochrome photochemistry in search of Pfr- favoured variants

Autores
Antelo, Giuliano Tomás; Sánchez-Lamas, Maximiliano; Goldbaum, Fernando Alberto; Otero, Lisandro Horacio; Bonomi, Hernán Ruy; Rinaldi, Jimena Julieta
Año de publicación
2020
Idioma
inglés
Tipo de recurso
documento de conferencia
Estado
versión publicada
Descripción
Phytochromes are photosensitive proteins with a covalently bound open-chain chromophore that can switch between two principal states: red-light absorbing Pr and far-red light absorbing Pfr. Our group has previously shown that the bacteriophytochrome from Xanthomonas campestris pv. campestris (XccBphP) is a bathy-like phytochrome that uses biliverdin IXα as a co-factor and is involved in bacterial virulence (Bonomi H.R. et al., 2016; Otero L.H. et al., 2016). To date, the XccBphP crystal structure could only be solved in the Pr state (Klinke S. et al., 2014; Otero L.H. et al., 2016), while the structure of its Pfr state remains elusive. The aims of this work were to develop an efficient screening methodology for the rapid characterization and to identify XccBphP variants that favour the Pfr form. The screening approach developed here consists in analyzing the UV-Vis absorption behaviour of clarified crude extracts containing recombinant phytochromes. This strategy has allowed us to quickly explore over a hundred XccBphP variants, characterize multiple variants and identify Pfr-favoured candidates. The high-quality data obtained enabled not only a qualitative, but also a quantitative characterization of their photochemistry. This method could be easily adapted to other phytochromes or other photoreceptor families.
Fil: Antelo, Giuliano Tomás. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; Argentina
Fil: Sánchez-Lamas, Maximiliano. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; Argentina
Fil: Goldbaum, Fernando Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; Argentina
Fil: Otero, Lisandro Horacio. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; Argentina
Fil: Bonomi, Hernán Ruy. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; Argentina
Fil: Rinaldi, Jimena Julieta. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; Argentina
V Reunión Grupo Argentino de Fotobiología
La Plata
Argentina
Grupo Argentino de Fotobiología
Materia
BACTERIOPHYTOCHROME
XANTHOMONAS
Fotorreceptores
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/154230
Acceder
id CONICETDig_7b86309f2fc756bcf922db859b67fce3
oai_identifier_str oai:ri.conicet.gov.ar:11336/154230
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling A spectroscopy-based methodology for rapid screening and characterization of phytochrome photochemistry in search of Pfr- favoured variantsAntelo, Giuliano TomásSánchez-Lamas, MaximilianoGoldbaum, Fernando AlbertoOtero, Lisandro HoracioBonomi, Hernán RuyRinaldi, Jimena JulietaBACTERIOPHYTOCHROMEXANTHOMONASFotorreceptoreshttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Phytochromes are photosensitive proteins with a covalently bound open-chain chromophore that can switch between two principal states: red-light absorbing Pr and far-red light absorbing Pfr. Our group has previously shown that the bacteriophytochrome from Xanthomonas campestris pv. campestris (XccBphP) is a bathy-like phytochrome that uses biliverdin IXα as a co-factor and is involved in bacterial virulence (Bonomi H.R. et al., 2016; Otero L.H. et al., 2016). To date, the XccBphP crystal structure could only be solved in the Pr state (Klinke S. et al., 2014; Otero L.H. et al., 2016), while the structure of its Pfr state remains elusive. The aims of this work were to develop an efficient screening methodology for the rapid characterization and to identify XccBphP variants that favour the Pfr form. The screening approach developed here consists in analyzing the UV-Vis absorption behaviour of clarified crude extracts containing recombinant phytochromes. This strategy has allowed us to quickly explore over a hundred XccBphP variants, characterize multiple variants and identify Pfr-favoured candidates. The high-quality data obtained enabled not only a qualitative, but also a quantitative characterization of their photochemistry. This method could be easily adapted to other phytochromes or other photoreceptor families.Fil: Antelo, Giuliano Tomás. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; ArgentinaFil: Sánchez-Lamas, Maximiliano. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; ArgentinaFil: Goldbaum, Fernando Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; ArgentinaFil: Otero, Lisandro Horacio. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; ArgentinaFil: Bonomi, Hernán Ruy. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; ArgentinaFil: Rinaldi, Jimena Julieta. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; ArgentinaV Reunión Grupo Argentino de FotobiologíaLa PlataArgentinaGrupo Argentino de FotobiologíaGrupo Argentino de Fotobiología2020info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/conferenceObjectReuniónBookhttp://purl.org/coar/resource_type/c_5794info:ar-repo/semantics/documentoDeConferenciaapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/154230A spectroscopy-based methodology for rapid screening and characterization of phytochrome photochemistry in search of Pfr- favoured variants; V Reunión Grupo Argentino de Fotobiología; La Plata; Argentina; 2020; 1-9CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://grupoargentinodefotobiologia.info/site/site/grupar/pluginfile.php/94/mod_page/content/99/VGRAFOBLibroderesumenes.pdfinfo:eu-repo/semantics/altIdentifier/url/https://grupoargentinodefotobiologia.info/site/site/grupar/mod/page/view.php?id=14Nacionalinfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-10T13:23:23Zoai:ri.conicet.gov.ar:11336/154230instacron: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:34982025-09-10 13:23:23.338CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv A spectroscopy-based methodology for rapid screening and characterization of phytochrome photochemistry in search of Pfr- favoured variants
title A spectroscopy-based methodology for rapid screening and characterization of phytochrome photochemistry in search of Pfr- favoured variants
spellingShingle A spectroscopy-based methodology for rapid screening and characterization of phytochrome photochemistry in search of Pfr- favoured variants
Antelo, Giuliano Tomás
BACTERIOPHYTOCHROME
XANTHOMONAS
Fotorreceptores
title_short A spectroscopy-based methodology for rapid screening and characterization of phytochrome photochemistry in search of Pfr- favoured variants
title_full A spectroscopy-based methodology for rapid screening and characterization of phytochrome photochemistry in search of Pfr- favoured variants
title_fullStr A spectroscopy-based methodology for rapid screening and characterization of phytochrome photochemistry in search of Pfr- favoured variants
title_full_unstemmed A spectroscopy-based methodology for rapid screening and characterization of phytochrome photochemistry in search of Pfr- favoured variants
title_sort A spectroscopy-based methodology for rapid screening and characterization of phytochrome photochemistry in search of Pfr- favoured variants
dc.creator.none.fl_str_mv Antelo, Giuliano Tomás
Sánchez-Lamas, Maximiliano
Goldbaum, Fernando Alberto
Otero, Lisandro Horacio
Bonomi, Hernán Ruy
Rinaldi, Jimena Julieta
author Antelo, Giuliano Tomás
author_facet Antelo, Giuliano Tomás
Sánchez-Lamas, Maximiliano
Goldbaum, Fernando Alberto
Otero, Lisandro Horacio
Bonomi, Hernán Ruy
Rinaldi, Jimena Julieta
author_role author
author2 Sánchez-Lamas, Maximiliano
Goldbaum, Fernando Alberto
Otero, Lisandro Horacio
Bonomi, Hernán Ruy
Rinaldi, Jimena Julieta
author2_role author
author
author
author
author
dc.subject.none.fl_str_mv BACTERIOPHYTOCHROME
XANTHOMONAS
Fotorreceptores
topic BACTERIOPHYTOCHROME
XANTHOMONAS
Fotorreceptores
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Phytochromes are photosensitive proteins with a covalently bound open-chain chromophore that can switch between two principal states: red-light absorbing Pr and far-red light absorbing Pfr. Our group has previously shown that the bacteriophytochrome from Xanthomonas campestris pv. campestris (XccBphP) is a bathy-like phytochrome that uses biliverdin IXα as a co-factor and is involved in bacterial virulence (Bonomi H.R. et al., 2016; Otero L.H. et al., 2016). To date, the XccBphP crystal structure could only be solved in the Pr state (Klinke S. et al., 2014; Otero L.H. et al., 2016), while the structure of its Pfr state remains elusive. The aims of this work were to develop an efficient screening methodology for the rapid characterization and to identify XccBphP variants that favour the Pfr form. The screening approach developed here consists in analyzing the UV-Vis absorption behaviour of clarified crude extracts containing recombinant phytochromes. This strategy has allowed us to quickly explore over a hundred XccBphP variants, characterize multiple variants and identify Pfr-favoured candidates. The high-quality data obtained enabled not only a qualitative, but also a quantitative characterization of their photochemistry. This method could be easily adapted to other phytochromes or other photoreceptor families.
Fil: Antelo, Giuliano Tomás. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; Argentina
Fil: Sánchez-Lamas, Maximiliano. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; Argentina
Fil: Goldbaum, Fernando Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; Argentina
Fil: Otero, Lisandro Horacio. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; Argentina
Fil: Bonomi, Hernán Ruy. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; Argentina
Fil: Rinaldi, Jimena Julieta. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; Argentina
V Reunión Grupo Argentino de Fotobiología
La Plata
Argentina
Grupo Argentino de Fotobiología
description Phytochromes are photosensitive proteins with a covalently bound open-chain chromophore that can switch between two principal states: red-light absorbing Pr and far-red light absorbing Pfr. Our group has previously shown that the bacteriophytochrome from Xanthomonas campestris pv. campestris (XccBphP) is a bathy-like phytochrome that uses biliverdin IXα as a co-factor and is involved in bacterial virulence (Bonomi H.R. et al., 2016; Otero L.H. et al., 2016). To date, the XccBphP crystal structure could only be solved in the Pr state (Klinke S. et al., 2014; Otero L.H. et al., 2016), while the structure of its Pfr state remains elusive. The aims of this work were to develop an efficient screening methodology for the rapid characterization and to identify XccBphP variants that favour the Pfr form. The screening approach developed here consists in analyzing the UV-Vis absorption behaviour of clarified crude extracts containing recombinant phytochromes. This strategy has allowed us to quickly explore over a hundred XccBphP variants, characterize multiple variants and identify Pfr-favoured candidates. The high-quality data obtained enabled not only a qualitative, but also a quantitative characterization of their photochemistry. This method could be easily adapted to other phytochromes or other photoreceptor families.
publishDate 2020
dc.date.none.fl_str_mv 2020
dc.type.none.fl_str_mv info:eu-repo/semantics/publishedVersion
info:eu-repo/semantics/conferenceObject
Reunión
Book
http://purl.org/coar/resource_type/c_5794
info:ar-repo/semantics/documentoDeConferencia
status_str publishedVersion
format conferenceObject
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/154230
A spectroscopy-based methodology for rapid screening and characterization of phytochrome photochemistry in search of Pfr- favoured variants; V Reunión Grupo Argentino de Fotobiología; La Plata; Argentina; 2020; 1-9
CONICET Digital
CONICET
url http://hdl.handle.net/11336/154230
identifier_str_mv A spectroscopy-based methodology for rapid screening and characterization of phytochrome photochemistry in search of Pfr- favoured variants; V Reunión Grupo Argentino de Fotobiología; La Plata; Argentina; 2020; 1-9
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://grupoargentinodefotobiologia.info/site/site/grupar/pluginfile.php/94/mod_page/content/99/VGRAFOBLibroderesumenes.pdf
info:eu-repo/semantics/altIdentifier/url/https://grupoargentinodefotobiologia.info/site/site/grupar/mod/page/view.php?id=14
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.coverage.none.fl_str_mv Nacional
dc.publisher.none.fl_str_mv Grupo Argentino de Fotobiología
publisher.none.fl_str_mv Grupo Argentino de Fotobiología
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_ 1842981291151065088
score 12.48226

Publicaciones similares