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
- Institución
- Consejo Nacional de Investigaciones Científicas y Técnicas
- OAI Identificador
- oai:ri.conicet.gov.ar:11336/83697
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CONICET Digital (CONICET) |
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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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1844613005795393536 |
score |
13.070432 |