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dc.contributor.author
Orihuela, Rubén
dc.contributor.author
Monteiro, Freddy
dc.contributor.author
Pagani, María Ayelén
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dc.contributor.author
Capdevila, Merce
dc.contributor.author
Atrian, Silvia
dc.date.available
2017-04-18T17:55:22Z
dc.date.issued
2010-11
dc.identifier.citation
Orihuela, Rubén; Monteiro, Freddy; Pagani, María Ayelén; Capdevila, Merce; Atrian, Silvia; Evidence of native Metal–S2–Metallothionein complexes confirmed by the analysis of Cup1 Divalent-Metal-Ion binding properties; Wiley VCH Verlag; Chemistry- A European Journal; 16; 41; 11-2010; 12363-12372
dc.identifier.uri
http://hdl.handle.net/11336/15375
dc.description.abstract
It has previously been shown that recombinant synthesis, under metal-supplemented conditions, of diverse metallothioneins (MTs) results in the recovery of a subpopulation of S2- containing complexes in addition to the S2-devoid canonical metal–MT species. Further significance of this finding has remained veiled by the possibility of it being a mere consequence of synthesis in a heterologous bacterial system. Herein, we present definitive evidence that S2 ligands are also constituents of native metal–MT complexes. Because, although practically universal, the highest S2 content is incorporated by copper-thioneins when coordinating divalent metal ions, we adapted the Saccharomyces cerevisiae Cup1 protein, which is the most paradigmatic copper-thionein, as an experimental model. Most significantly, native Cd–Cup1 complexes were purified and fully spectroscopically and spectrometrically characterized from the 301N mutant yeast strain, which allows Cup1 synthesis even in the absence of copper. These results undoubtedly revealed the presence of a Cd– S2–Cup1 species in native preparations, which were only recovered when carefully avoiding the use of ion-exchange chromatography in the purification protocol. Furthermore, complete analysis of recombinant (Escherichia coli) Zn–Cup1, Cd–Cup1, and Cu– Cup1 and those complexes that result from Zn/Cd and Zn/Cu replacements in vitro and acidification/renaturalization processes yielded a comprehensive and comparative overview of the metal-binding abilities of Cup1. Overall, we consider the main conclusions of this study to go beyond the mere study of the particular Cup1 MT, so that they should be considered to delineate a new point of view on the interaction between copper-thioneins and divalent metal ions, still an unexplored aspect in MT research.
dc.format
application/pdf
dc.language.iso
eng
dc.publisher
Wiley VCH Verlag
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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
Bioinorganic Chemistry
dc.subject
Circular Dichroism
dc.subject
Metalloproteins
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Metallothioneins
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Sulfide Ligands
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Otras Ciencias Químicas
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dc.subject.classification
Ciencias Químicas
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dc.subject.classification
CIENCIAS NATURALES Y EXACTAS
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dc.subject.classification
Bioquímica y Biología Molecular
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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
Evidence of native Metal–S2–Metallothionein complexes confirmed by the analysis of Cup1 Divalent-Metal-Ion binding properties
dc.type
info:eu-repo/semantics/article
dc.type
info:ar-repo/semantics/artículo
dc.type
info:eu-repo/semantics/publishedVersion
dc.date.updated
2017-04-18T14:02:16Z
dc.identifier.eissn
1521-3765
dc.journal.volume
16
dc.journal.number
41
dc.journal.pagination
12363-12372
dc.journal.pais
Alemania
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dc.journal.ciudad
Weinheim
dc.description.fil
Fil: Orihuela, Rubén. Universitat Autonoma de Barcelona; España
dc.description.fil
Fil: Monteiro, Freddy. Universidad de Barcelona; España
dc.description.fil
Fil: Pagani, María Ayelén. Universidad de Barcelona; España. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Rosario. Centro de Estudios Fotosintéticos y Bioquímicos (i); Argentina. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas; Argentina
dc.description.fil
Fil: Capdevila, Merce. Universitat Autonoma de Barcelona; España
dc.description.fil
Fil: Atrian, Silvia. Universidad de Barcelona; España
dc.journal.title
Chemistry- A European Journal
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dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/doi/http://dx.doi.org/10.1002/chem.201001125
dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/url/http://onlinelibrary.wiley.com/doi/10.1002/chem.201001125/abstract
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