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dc.contributor.author
Kraiselburd, Ivana
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dc.contributor.author
Gutt, Alexander
dc.contributor.author
Losi, Aba
dc.contributor.author
Gärtner, Wolfgang
dc.contributor.author
Orellano, Elena Graciela
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dc.date.available
2018-07-23T17:23:15Z
dc.date.issued
2015-09
dc.identifier.citation
Kraiselburd, Ivana; Gutt, Alexander; Losi, Aba; Gärtner, Wolfgang; Orellano, Elena Graciela; Functional Characterization of a LOV-Histidine Kinase Photoreceptor from Xanthomonas citri subsp. citri; Wiley Blackwell Publishing, Inc; Photochemistry and Photobiology; 91; 5; 9-2015; 1123-1132
dc.identifier.issn
0031-8655
dc.identifier.uri
http://hdl.handle.net/11336/52828
dc.description.abstract
The blue-light (BL) absorbing protein Xcc-LOV from Xanthomonas citri subsp. citri is composed of a LOV-domain, a histidine kinase (HK) and a response regulator. Spectroscopic characterization of Xcc-LOV identified intermediates and kinetics of the protein's photocycle. Measurements of steady state and time-resolved fluorescence allowed determination of quantum yields for triplet (ΦT = 0.68 ± 0.03) and photoproduct formation (Φ390 = 0.46 ± 0.05). The lifetime for triplet decay was determined as τT = 2.4-2.8 μs. Fluorescence of tryptophan and tyrosine residues was unchanged upon light-to-dark conversion, emphasizing the absence of significant conformational changes. Photochemistry was blocked upon cysteine C76 (C76S) mutation, causing a seven-fold longer lifetime of the triplet state (τT = 16-18.5 μs). Optoacoustic spectroscopy yielded the energy content of the triplet state. Interestingly, Xcc-LOV did not undergo the volume contraction reported for other LOV domains within the observation time window, although the back-conversion into the dark state was accompanied by a volume expansion. A radioactivity-based enzyme function assay revealed a larger HK activity in the lit than in the dark state. The C76S mutant showed a still lower enzyme function, indicating the dark state activity being corrupted by a remaining portion of the long-lived lit state.
dc.format
application/pdf
dc.language.iso
eng
dc.publisher
Wiley Blackwell Publishing, Inc
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dc.rights
info:eu-repo/semantics/openAccess
dc.rights.uri
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.subject
Lov Protein
dc.subject
Histidine Kinase
dc.subject
Flash Photolysis
dc.subject
Optoacustics
dc.subject.classification
Otras Ciencias Biológicas
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dc.subject.classification
Ciencias Biológicas
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dc.subject.classification
CIENCIAS NATURALES Y EXACTAS
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dc.title
Functional Characterization of a LOV-Histidine Kinase Photoreceptor from Xanthomonas citri subsp. citri
dc.type
info:eu-repo/semantics/article
dc.type
info:ar-repo/semantics/artículo
dc.type
info:eu-repo/semantics/publishedVersion
dc.date.updated
2018-07-20T18:03:02Z
dc.journal.volume
91
dc.journal.number
5
dc.journal.pagination
1123-1132
dc.journal.pais
Reino Unido
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dc.journal.ciudad
Londres
dc.description.fil
Fil: Kraiselburd, Ivana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Biología Molecular y Celular de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Biología Molecular y Celular de Rosario; Argentina
dc.description.fil
Fil: Gutt, Alexander. Max‐Planck‐Institute for Chemical Energy Conversion; Alemania
dc.description.fil
Fil: Losi, Aba. Università di Parma; Italia
dc.description.fil
Fil: Gärtner, Wolfgang. Max‐Planck‐Institute for Chemical Energy Conversion; Alemania
dc.description.fil
Fil: Orellano, Elena Graciela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Biología Molecular y Celular de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Biología Molecular y Celular de Rosario; Argentina
dc.journal.title
Photochemistry and Photobiology
dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/doi/https://dx.doi.org/10.1111/php.12493
dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/url/https://onlinelibrary.wiley.com/doi/abs/10.1111/php.12493
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