Hints for Metal-Preference Protein Sequence Determinants: Different Metal Binding Features of the Five Tetrahymena thermophila Metallothioneins

The metal binding preference of metallothioneins (MTs) groups them in two extreme subsets, the Zn/Cd - and the Cu - thioneins. Ciliates harbor the largest MT gene/protein family reported so far, i n- cluding 5 paralogs that exhibit relatively low sequence similarity, excepting MTT2 and MTT4. I n Te...

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Autores: Espart Herrero, Anna, Marín, Maribel, Gil Moreno, Selene, Palacios Bonilla, Òscar, Amaro, Francisco, Martín González, Ana, Gutiérrez, Juan C., Capdevila Vidal, Mercè, Atrian i Ventura, Sílvia
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2015
País:España
Institución:Varias* (Consorci de Biblioteques Universitáries de Catalunya, Centre de Serveis Científics i Acadèmics de Catalunya)
Repositorio:Recercat. Dipósit de la Recerca de Catalunya
OAI Identifier:oai:recercat.cat:2445/149450
Acceso en línea:https://hdl.handle.net/2445/149450
Access Level:acceso abierto
Palabra clave:Proteïnes
Metal·loproteïnes
Proteins
Metalloproteins
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spelling Hints for Metal-Preference Protein Sequence Determinants: Different Metal Binding Features of the Five Tetrahymena thermophila MetallothioneinsEspart Herrero, AnnaMarín, MaribelGil Moreno, SelenePalacios Bonilla, ÒscarAmaro, FranciscoMartín González, AnaGutiérrez, Juan C.Capdevila Vidal, MercèAtrian i Ventura, SílviaProteïnesMetal·loproteïnesProteinsMetalloproteinsThe metal binding preference of metallothioneins (MTs) groups them in two extreme subsets, the Zn/Cd - and the Cu - thioneins. Ciliates harbor the largest MT gene/protein family reported so far, i n- cluding 5 paralogs that exhibit relatively low sequence similarity, excepting MTT2 and MTT4. I n Te t- rahymena thermophila , three MTs (MTT1, MTT3 and MTT5) were considered Cd - thioneins and two (MTT2 and MTT4) Cu - thioneins, according to gene expression inducibility and phylogenetic analysis. In this study, the metal - binding abilities of the five MTT pr oteins were characterized, to obtain information about the folding and stability of their cognate - and non - cognate metal complexes, and to characterize the T. thermophila MT system at protein level. Hence, the five MTTs were recombinantly synthesized as Zn 2+ - , Cd 2+ - or Cu + - complexes, which were analyzed by electrospray mass spectrometry (ESI - MS), circular dichroism (CD), and UV - vis spectrophotometry. Among the Cd - thioneins, MTT1 and MTT5 were optimal for Cd 2+ coordination, yielding unique Cd 17 - and Cd 8 - com plexes, respectively. When binding Zn 2+ , they rendered a mixture of Zn - species. Only MTT5 was capable to coordinate Cu + , although yielding heteronuclear Zn - , Cu - species or highly unstable Cu - homometallic species. MTT3 exhibited poor binding abilities both for Cd 2+ and for Cu + , and although not optimally, it yielded the best result when coordinating Zn 2+ . The two Cu - thioneins, MTT2 and MTT4 isoforms formed homometallic Cu - complexes (major Cu 20 - MTT) upon synthesis in Cu - supplemented hosts. Contrarily, they we re unable to fold into stable Cd - complexes, while Zn - MTT species were only recovered for MTT4 (major Zn 10 - MTT4). Thus, the metal binding preferences of the five T. thermophila MTs correlate well with their previous classification as Cd - and Cu - thioneins, a nd globally, they can be classified from Zn/Cd - to Cu - thioneins according to the gradation: MTT1>MTT5>MTT3>MTT4>MTT2. The main mechanisms underlying the evolution and specialization of the MTT metal binding preferences may have been i n- ternal tandem duplica tions, presence of doublet and triplet Cys patterns in Zn/Cd - thioneins, and o p- timization of site specific amino acid determinants (Lys for Zn/Cd - and Asn for Cu - coordination).Ivyspring International2020202020152020info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersion16 p.application/pdfhttps://hdl.handle.net/2445/149450Articles publicats en revistes (Genètica, Microbiologia i Estadística)reponame:Recercat. Dipósit de la Recerca de Catalunyainstname:Varias* (Consorci de Biblioteques Universitáries de Catalunya, Centre de Serveis Científics i Acadèmics de Catalunya)InglésReproducció del document publicat a: https://doi.org/10.7150/ijbs.11060International Journal of Biological Sciences, 2015, vol. 11, num. 4, p. 456-471https://doi.org/10.7150/ijbs.11060cc-by-nc-nd (c) Ivyspring International, 2015http://creativecommons.org/licenses/by-nc-nd/3.0/esinfo:eu-repo/semantics/openAccessoai:recercat.cat:2445/1494502026-05-29T05:05:01Z
dc.title.none.fl_str_mv Hints for Metal-Preference Protein Sequence Determinants: Different Metal Binding Features of the Five Tetrahymena thermophila Metallothioneins
title Hints for Metal-Preference Protein Sequence Determinants: Different Metal Binding Features of the Five Tetrahymena thermophila Metallothioneins
spellingShingle Hints for Metal-Preference Protein Sequence Determinants: Different Metal Binding Features of the Five Tetrahymena thermophila Metallothioneins
Espart Herrero, Anna
Proteïnes
Metal·loproteïnes
Proteins
Metalloproteins
title_short Hints for Metal-Preference Protein Sequence Determinants: Different Metal Binding Features of the Five Tetrahymena thermophila Metallothioneins
title_full Hints for Metal-Preference Protein Sequence Determinants: Different Metal Binding Features of the Five Tetrahymena thermophila Metallothioneins
title_fullStr Hints for Metal-Preference Protein Sequence Determinants: Different Metal Binding Features of the Five Tetrahymena thermophila Metallothioneins
title_full_unstemmed Hints for Metal-Preference Protein Sequence Determinants: Different Metal Binding Features of the Five Tetrahymena thermophila Metallothioneins
title_sort Hints for Metal-Preference Protein Sequence Determinants: Different Metal Binding Features of the Five Tetrahymena thermophila Metallothioneins
dc.creator.none.fl_str_mv Espart Herrero, Anna
Marín, Maribel
Gil Moreno, Selene
Palacios Bonilla, Òscar
Amaro, Francisco
Martín González, Ana
Gutiérrez, Juan C.
Capdevila Vidal, Mercè
Atrian i Ventura, Sílvia
author Espart Herrero, Anna
author_facet Espart Herrero, Anna
Marín, Maribel
Gil Moreno, Selene
Palacios Bonilla, Òscar
Amaro, Francisco
Martín González, Ana
Gutiérrez, Juan C.
Capdevila Vidal, Mercè
Atrian i Ventura, Sílvia
author_role author
author2 Marín, Maribel
Gil Moreno, Selene
Palacios Bonilla, Òscar
Amaro, Francisco
Martín González, Ana
Gutiérrez, Juan C.
Capdevila Vidal, Mercè
Atrian i Ventura, Sílvia
author2_role author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv Proteïnes
Metal·loproteïnes
Proteins
Metalloproteins
topic Proteïnes
Metal·loproteïnes
Proteins
Metalloproteins
description The metal binding preference of metallothioneins (MTs) groups them in two extreme subsets, the Zn/Cd - and the Cu - thioneins. Ciliates harbor the largest MT gene/protein family reported so far, i n- cluding 5 paralogs that exhibit relatively low sequence similarity, excepting MTT2 and MTT4. I n Te t- rahymena thermophila , three MTs (MTT1, MTT3 and MTT5) were considered Cd - thioneins and two (MTT2 and MTT4) Cu - thioneins, according to gene expression inducibility and phylogenetic analysis. In this study, the metal - binding abilities of the five MTT pr oteins were characterized, to obtain information about the folding and stability of their cognate - and non - cognate metal complexes, and to characterize the T. thermophila MT system at protein level. Hence, the five MTTs were recombinantly synthesized as Zn 2+ - , Cd 2+ - or Cu + - complexes, which were analyzed by electrospray mass spectrometry (ESI - MS), circular dichroism (CD), and UV - vis spectrophotometry. Among the Cd - thioneins, MTT1 and MTT5 were optimal for Cd 2+ coordination, yielding unique Cd 17 - and Cd 8 - com plexes, respectively. When binding Zn 2+ , they rendered a mixture of Zn - species. Only MTT5 was capable to coordinate Cu + , although yielding heteronuclear Zn - , Cu - species or highly unstable Cu - homometallic species. MTT3 exhibited poor binding abilities both for Cd 2+ and for Cu + , and although not optimally, it yielded the best result when coordinating Zn 2+ . The two Cu - thioneins, MTT2 and MTT4 isoforms formed homometallic Cu - complexes (major Cu 20 - MTT) upon synthesis in Cu - supplemented hosts. Contrarily, they we re unable to fold into stable Cd - complexes, while Zn - MTT species were only recovered for MTT4 (major Zn 10 - MTT4). Thus, the metal binding preferences of the five T. thermophila MTs correlate well with their previous classification as Cd - and Cu - thioneins, a nd globally, they can be classified from Zn/Cd - to Cu - thioneins according to the gradation: MTT1>MTT5>MTT3>MTT4>MTT2. The main mechanisms underlying the evolution and specialization of the MTT metal binding preferences may have been i n- ternal tandem duplica tions, presence of doublet and triplet Cys patterns in Zn/Cd - thioneins, and o p- timization of site specific amino acid determinants (Lys for Zn/Cd - and Asn for Cu - coordination).
publishDate 2015
dc.date.none.fl_str_mv 2015
2020
2020
2020
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv https://hdl.handle.net/2445/149450
url https://hdl.handle.net/2445/149450
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv Reproducció del document publicat a: https://doi.org/10.7150/ijbs.11060
International Journal of Biological Sciences, 2015, vol. 11, num. 4, p. 456-471
https://doi.org/10.7150/ijbs.11060
dc.rights.none.fl_str_mv cc-by-nc-nd (c) Ivyspring International, 2015
http://creativecommons.org/licenses/by-nc-nd/3.0/es
info:eu-repo/semantics/openAccess
rights_invalid_str_mv cc-by-nc-nd (c) Ivyspring International, 2015
http://creativecommons.org/licenses/by-nc-nd/3.0/es
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv 16 p.
application/pdf
dc.publisher.none.fl_str_mv Ivyspring International
publisher.none.fl_str_mv Ivyspring International
dc.source.none.fl_str_mv Articles publicats en revistes (Genètica, Microbiologia i Estadística)
reponame:Recercat. Dipósit de la Recerca de Catalunya
instname:Varias* (Consorci de Biblioteques Universitáries de Catalunya, Centre de Serveis Científics i Acadèmics de Catalunya)
instname_str Varias* (Consorci de Biblioteques Universitáries de Catalunya, Centre de Serveis Científics i Acadèmics de Catalunya)
reponame_str Recercat. Dipósit de la Recerca de Catalunya
collection Recercat. Dipósit de la Recerca de Catalunya
repository.name.fl_str_mv
repository.mail.fl_str_mv
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score 15,81155

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