Mutational analysis of Deinococcus radiodurans bacteriophytochrome reveals key amino acids necessary for the photochromicity and proton exchange cycle of phytochromes

Autores
Wagner, Jeremiah R.; Zhang, Junrui; Von Stetten, David; Günther, Mina; Murgida, Daniel Horacio; Mroginski, Maria Andrea; Walker, Joseph M.; Forest, Katrina T.; Hildebrandt, Peter; Vierstra, Richard D.
Año de publicación
2008
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
The ability of phytochromes (Phy) to act as photointerconvertible light switches in plants and microorganisms depends on key interactions between the bilin chromophore and the apoprotein that promote bilin attachment and photointerconversion between the spectrally distinct red light-absorbing Pr conformer and far red light-absorbing Pfr conformer. Using structurally guided site-directed mutagenesis combined with several spectroscopic methods, we examined the roles of conserved amino acids within the bilin-binding domain of Deinococcus radiodurans bacteriophytochrome with respect to chromophore ligation and Pr/Pfr photoconversion. Incorporation of biliverdin IXα (BV), its structure in the Pr state, and its ability to photoisomerize to the first photocycle intermediate are insensitive to most single mutations, implying that these properties are robust with respect to small structural/electrostatic alterations in the binding pocket. In contrast, photoconversion to Pfr is highly sensitive to the chromophore environment. Many of the variants form spectrally bleached Meta-type intermediates in red light that do not relax to Pfr. Particularly important are Asp-207 and His-260, which are invariant within the Phy superfamily and participate in a unique hydrogen bond matrix involving the A, B, and C pyrrole ring nitrogens of BV and their associated pyrrole water. Resonance Raman spectroscopy demonstrates that substitutions of these residues disrupt the Pr to Pfr protonation cycle of BV with the chromophore locked in a deprotonated Meta-Rc-like photoconversion intermediate after red light irradiation. Collectively, the data show that a number of contacts contribute to the unique photochromicity of Phy-type photoreceptors. These include residues that fix the bilin in the pocket, coordinate the pyrrole water, and possibly promote the proton exchange cycle during photoconversion.
Fil: Wagner, Jeremiah R.. Beloit College; Estados Unidos. University of Wisconsin; Estados Unidos
Fil: Zhang, Junrui. University of Wisconsin; Estados Unidos
Fil: Von Stetten, David. Technishe Universitat Berlin; Alemania
Fil: Günther, Mina. Technishe Universitat Berlin; Alemania
Fil: Murgida, Daniel Horacio. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Technishe Universitat Berlin; Alemania
Fil: Mroginski, Maria Andrea. Technishe Universitat Berlin; Alemania
Fil: Walker, Joseph M.. University of Wisconsin; Estados Unidos
Fil: Forest, Katrina T.. University of Wisconsin; Estados Unidos
Fil: Hildebrandt, Peter. Technishe Universitat Berlin; Alemania
Fil: Vierstra, Richard D.. University of Wisconsin; Estados Unidos
Materia
Phytochromes
Photoreceptors
FT-Raman
DFT calculations
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:11336/83697

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network_name_str CONICET Digital (CONICET)
spelling Mutational analysis of Deinococcus radiodurans bacteriophytochrome reveals key amino acids necessary for the photochromicity and proton exchange cycle of phytochromesWagner, Jeremiah R.Zhang, JunruiVon Stetten, DavidGünther, MinaMurgida, Daniel HoracioMroginski, Maria AndreaWalker, Joseph M.Forest, Katrina T.Hildebrandt, PeterVierstra, Richard D.PhytochromesPhotoreceptorsFT-RamanDFT calculationshttps://purl.org/becyt/ford/1.4https://purl.org/becyt/ford/1The ability of phytochromes (Phy) to act as photointerconvertible light switches in plants and microorganisms depends on key interactions between the bilin chromophore and the apoprotein that promote bilin attachment and photointerconversion between the spectrally distinct red light-absorbing Pr conformer and far red light-absorbing Pfr conformer. Using structurally guided site-directed mutagenesis combined with several spectroscopic methods, we examined the roles of conserved amino acids within the bilin-binding domain of Deinococcus radiodurans bacteriophytochrome with respect to chromophore ligation and Pr/Pfr photoconversion. Incorporation of biliverdin IXα (BV), its structure in the Pr state, and its ability to photoisomerize to the first photocycle intermediate are insensitive to most single mutations, implying that these properties are robust with respect to small structural/electrostatic alterations in the binding pocket. In contrast, photoconversion to Pfr is highly sensitive to the chromophore environment. Many of the variants form spectrally bleached Meta-type intermediates in red light that do not relax to Pfr. Particularly important are Asp-207 and His-260, which are invariant within the Phy superfamily and participate in a unique hydrogen bond matrix involving the A, B, and C pyrrole ring nitrogens of BV and their associated pyrrole water. Resonance Raman spectroscopy demonstrates that substitutions of these residues disrupt the Pr to Pfr protonation cycle of BV with the chromophore locked in a deprotonated Meta-Rc-like photoconversion intermediate after red light irradiation. Collectively, the data show that a number of contacts contribute to the unique photochromicity of Phy-type photoreceptors. These include residues that fix the bilin in the pocket, coordinate the pyrrole water, and possibly promote the proton exchange cycle during photoconversion.Fil: Wagner, Jeremiah R.. Beloit College; Estados Unidos. University of Wisconsin; Estados UnidosFil: Zhang, Junrui. University of Wisconsin; Estados UnidosFil: Von Stetten, David. Technishe Universitat Berlin; AlemaniaFil: Günther, Mina. Technishe Universitat Berlin; AlemaniaFil: Murgida, Daniel Horacio. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Technishe Universitat Berlin; AlemaniaFil: Mroginski, Maria Andrea. Technishe Universitat Berlin; AlemaniaFil: Walker, Joseph M.. University of Wisconsin; Estados UnidosFil: Forest, Katrina T.. University of Wisconsin; Estados UnidosFil: Hildebrandt, Peter. Technishe Universitat Berlin; AlemaniaFil: Vierstra, Richard D.. University of Wisconsin; Estados UnidosAmerican Society for Biochemistry and Molecular Biology2008-05info: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/11336/83697Wagner, Jeremiah R.; Zhang, Junrui; Von Stetten, David; Günther, Mina; Murgida, Daniel Horacio; et al.; Mutational analysis of Deinococcus radiodurans bacteriophytochrome reveals key amino acids necessary for the photochromicity and proton exchange cycle of phytochromes; American Society for Biochemistry and Molecular Biology; Journal of Biological Chemistry (online); 283; 18; 5-2008; 12212-122260021-9258CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.1074/jbc.M709355200info:eu-repo/semantics/altIdentifier/url/http://www.jbc.org/content/283/18/12212info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T09:32:52Zoai:ri.conicet.gov.ar:11336/83697instacron: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 09:32:52.989CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Mutational analysis of Deinococcus radiodurans bacteriophytochrome reveals key amino acids necessary for the photochromicity and proton exchange cycle of phytochromes
title Mutational analysis of Deinococcus radiodurans bacteriophytochrome reveals key amino acids necessary for the photochromicity and proton exchange cycle of phytochromes
spellingShingle Mutational analysis of Deinococcus radiodurans bacteriophytochrome reveals key amino acids necessary for the photochromicity and proton exchange cycle of phytochromes
Wagner, Jeremiah R.
Phytochromes
Photoreceptors
FT-Raman
DFT calculations
title_short Mutational analysis of Deinococcus radiodurans bacteriophytochrome reveals key amino acids necessary for the photochromicity and proton exchange cycle of phytochromes
title_full Mutational analysis of Deinococcus radiodurans bacteriophytochrome reveals key amino acids necessary for the photochromicity and proton exchange cycle of phytochromes
title_fullStr Mutational analysis of Deinococcus radiodurans bacteriophytochrome reveals key amino acids necessary for the photochromicity and proton exchange cycle of phytochromes
title_full_unstemmed Mutational analysis of Deinococcus radiodurans bacteriophytochrome reveals key amino acids necessary for the photochromicity and proton exchange cycle of phytochromes
title_sort Mutational analysis of Deinococcus radiodurans bacteriophytochrome reveals key amino acids necessary for the photochromicity and proton exchange cycle of phytochromes
dc.creator.none.fl_str_mv Wagner, Jeremiah R.
Zhang, Junrui
Von Stetten, David
Günther, Mina
Murgida, Daniel Horacio
Mroginski, Maria Andrea
Walker, Joseph M.
Forest, Katrina T.
Hildebrandt, Peter
Vierstra, Richard D.
author Wagner, Jeremiah R.
author_facet Wagner, Jeremiah R.
Zhang, Junrui
Von Stetten, David
Günther, Mina
Murgida, Daniel Horacio
Mroginski, Maria Andrea
Walker, Joseph M.
Forest, Katrina T.
Hildebrandt, Peter
Vierstra, Richard D.
author_role author
author2 Zhang, Junrui
Von Stetten, David
Günther, Mina
Murgida, Daniel Horacio
Mroginski, Maria Andrea
Walker, Joseph M.
Forest, Katrina T.
Hildebrandt, Peter
Vierstra, Richard D.
author2_role author
author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv Phytochromes
Photoreceptors
FT-Raman
DFT calculations
topic Phytochromes
Photoreceptors
FT-Raman
DFT calculations
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.4
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv The ability of phytochromes (Phy) to act as photointerconvertible light switches in plants and microorganisms depends on key interactions between the bilin chromophore and the apoprotein that promote bilin attachment and photointerconversion between the spectrally distinct red light-absorbing Pr conformer and far red light-absorbing Pfr conformer. Using structurally guided site-directed mutagenesis combined with several spectroscopic methods, we examined the roles of conserved amino acids within the bilin-binding domain of Deinococcus radiodurans bacteriophytochrome with respect to chromophore ligation and Pr/Pfr photoconversion. Incorporation of biliverdin IXα (BV), its structure in the Pr state, and its ability to photoisomerize to the first photocycle intermediate are insensitive to most single mutations, implying that these properties are robust with respect to small structural/electrostatic alterations in the binding pocket. In contrast, photoconversion to Pfr is highly sensitive to the chromophore environment. Many of the variants form spectrally bleached Meta-type intermediates in red light that do not relax to Pfr. Particularly important are Asp-207 and His-260, which are invariant within the Phy superfamily and participate in a unique hydrogen bond matrix involving the A, B, and C pyrrole ring nitrogens of BV and their associated pyrrole water. Resonance Raman spectroscopy demonstrates that substitutions of these residues disrupt the Pr to Pfr protonation cycle of BV with the chromophore locked in a deprotonated Meta-Rc-like photoconversion intermediate after red light irradiation. Collectively, the data show that a number of contacts contribute to the unique photochromicity of Phy-type photoreceptors. These include residues that fix the bilin in the pocket, coordinate the pyrrole water, and possibly promote the proton exchange cycle during photoconversion.
Fil: Wagner, Jeremiah R.. Beloit College; Estados Unidos. University of Wisconsin; Estados Unidos
Fil: Zhang, Junrui. University of Wisconsin; Estados Unidos
Fil: Von Stetten, David. Technishe Universitat Berlin; Alemania
Fil: Günther, Mina. Technishe Universitat Berlin; Alemania
Fil: Murgida, Daniel Horacio. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Technishe Universitat Berlin; Alemania
Fil: Mroginski, Maria Andrea. Technishe Universitat Berlin; Alemania
Fil: Walker, Joseph M.. University of Wisconsin; Estados Unidos
Fil: Forest, Katrina T.. University of Wisconsin; Estados Unidos
Fil: Hildebrandt, Peter. Technishe Universitat Berlin; Alemania
Fil: Vierstra, Richard D.. University of Wisconsin; Estados Unidos
description The ability of phytochromes (Phy) to act as photointerconvertible light switches in plants and microorganisms depends on key interactions between the bilin chromophore and the apoprotein that promote bilin attachment and photointerconversion between the spectrally distinct red light-absorbing Pr conformer and far red light-absorbing Pfr conformer. Using structurally guided site-directed mutagenesis combined with several spectroscopic methods, we examined the roles of conserved amino acids within the bilin-binding domain of Deinococcus radiodurans bacteriophytochrome with respect to chromophore ligation and Pr/Pfr photoconversion. Incorporation of biliverdin IXα (BV), its structure in the Pr state, and its ability to photoisomerize to the first photocycle intermediate are insensitive to most single mutations, implying that these properties are robust with respect to small structural/electrostatic alterations in the binding pocket. In contrast, photoconversion to Pfr is highly sensitive to the chromophore environment. Many of the variants form spectrally bleached Meta-type intermediates in red light that do not relax to Pfr. Particularly important are Asp-207 and His-260, which are invariant within the Phy superfamily and participate in a unique hydrogen bond matrix involving the A, B, and C pyrrole ring nitrogens of BV and their associated pyrrole water. Resonance Raman spectroscopy demonstrates that substitutions of these residues disrupt the Pr to Pfr protonation cycle of BV with the chromophore locked in a deprotonated Meta-Rc-like photoconversion intermediate after red light irradiation. Collectively, the data show that a number of contacts contribute to the unique photochromicity of Phy-type photoreceptors. These include residues that fix the bilin in the pocket, coordinate the pyrrole water, and possibly promote the proton exchange cycle during photoconversion.
publishDate 2008
dc.date.none.fl_str_mv 2008-05
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/11336/83697
Wagner, Jeremiah R.; Zhang, Junrui; Von Stetten, David; Günther, Mina; Murgida, Daniel Horacio; et al.; Mutational analysis of Deinococcus radiodurans bacteriophytochrome reveals key amino acids necessary for the photochromicity and proton exchange cycle of phytochromes; American Society for Biochemistry and Molecular Biology; Journal of Biological Chemistry (online); 283; 18; 5-2008; 12212-12226
0021-9258
CONICET Digital
CONICET
url http://hdl.handle.net/11336/83697
identifier_str_mv Wagner, Jeremiah R.; Zhang, Junrui; Von Stetten, David; Günther, Mina; Murgida, Daniel Horacio; et al.; Mutational analysis of Deinococcus radiodurans bacteriophytochrome reveals key amino acids necessary for the photochromicity and proton exchange cycle of phytochromes; American Society for Biochemistry and Molecular Biology; Journal of Biological Chemistry (online); 283; 18; 5-2008; 12212-12226
0021-9258
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/10.1074/jbc.M709355200
info:eu-repo/semantics/altIdentifier/url/http://www.jbc.org/content/283/18/12212
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
dc.publisher.none.fl_str_mv American Society for Biochemistry and Molecular Biology
publisher.none.fl_str_mv American Society for Biochemistry and Molecular Biology
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
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