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

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dc.contributor.authorEspart, Anna
dc.contributor.authorMarín, Maribel
dc.contributor.authorGil Moreno, Selene
dc.contributor.authorPalacios, Óscar
dc.contributor.authorAmaro Torres, Francisco
dc.contributor.authorMartín González, Ana María
dc.contributor.authorGutiérrez Fernández, Juan Carlos
dc.contributor.authorCapdevilla, Mercé
dc.contributor.authorAtrian, Sílvia
dc.date.accessioned2023-06-19T15:09:24Z
dc.date.available2023-06-19T15:09:24Z
dc.date.issued2015
dc.description.abstractThe 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, in-cluding 5 paralogs that exhibit relatively low sequence similarity, excepting MTT2 and MTT4. In Tet-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 proteins 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 Zn2+-, Cd2+- 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 Cd2+ coordination, yielding unique Cd17- and Cd8- complexes, respectively. When binding Zn2+, 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 Cd2+ and for Cu+, and although not optimally, it yielded the best result when coordinating Zn2+. The two Cu-thioneins, MTT2 and MTT4 isoforms formed homometallic Cu-complexes (major Cu20-MTT) upon synthesis in Cu-supplemented hosts. Contrarily, they were unable to fold into stable Cd-complexes, while Zn-MTT species were only recovered for MTT4 (major Zn10-MTT4). Thus, the metal binding preferences of the five T. thermophila MTs correlate well with their previous classification as Cd- and Cu-thioneins, and 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 in-ternal tandem duplications, presence of doublet and triplet Cys patterns in Zn/Cd-thioneins, and op-timization of site specific amino acid determinants (Lys for Zn/Cd- and Asn for Cu-coordination).
dc.description.departmentDepto. de Genética, Fisiología y Microbiología
dc.description.facultyFac. de Ciencias Biológicas
dc.description.refereedTRUE
dc.description.sponsorshipMinisterio de Economía y Competitividad (MINECO)
dc.description.statuspub
dc.eprint.idhttps://eprints.ucm.es/id/eprint/42342
dc.identifier.doi10.7150/ijbs.11060
dc.identifier.issn1449-2288
dc.identifier.officialurlhttp://www.ijbs.com/
dc.identifier.urihttps://hdl.handle.net/20.500.14352/35448
dc.issue.number4
dc.journal.titleInternational Journal of Biological Sciences
dc.language.isoeng
dc.page.final471
dc.page.initial456
dc.publisherIvyspring International Publisher
dc.relation.projectIDBIO2012-39682-C02-01
dc.relation.projectIDBIO2012-39682-C02-02
dc.relation.projectIDCGL2008-00317/BOS
dc.relation.projectIDBES-2010-036553
dc.rightsAtribución 3.0 España
dc.rights.accessRightsopen access
dc.rights.urihttps://creativecommons.org/licenses/by/3.0/es/
dc.subject.cdu579
dc.subject.cdu593.1
dc.subject.keywordMetallothionein
dc.subject.keywordFunctional Differentiation
dc.subject.keywordMetal specificity
dc.subject.keywordZinc
dc.subject.keywordCopper
dc.subject.keywordTetrahymena thermophila.
dc.subject.ucmInvertebrados
dc.subject.ucmMicrobiología (Biología)
dc.subject.unesco2401.17 Invertebrados
dc.subject.unesco2414 Microbiología
dc.titleHints for Metal-Preference Protein Sequence Determinants: Different Metal Binding Features of the Five Tetrahymena thermophila Metallothioneins
dc.typejournal article
dc.volume.number11
dspace.entity.typePublication
relation.isAuthorOfPublication9517917a-13ff-409d-b08f-bd204a61d258
relation.isAuthorOfPublicatione48abb18-103b-4edc-9081-81a44bc92b44
relation.isAuthorOfPublication0942b309-8e93-4ae5-a284-d49cb191c64b
relation.isAuthorOfPublication.latestForDiscovery9517917a-13ff-409d-b08f-bd204a61d258
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