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dc.contributorUniversitat Ramon Llull. IQS
dc.contributor.authorPlanas, Antoni (Planas Sauter)
dc.contributor.authorArsequell, Gemma
dc.contributor.authorCotrina, Ellen Y.
dc.contributor.authorVilà, Marta
dc.contributor.authorNieto, Joan
dc.date.accessioned2022-03-07T16:33:45Z
dc.date.accessioned2023-07-13T05:43:53Z
dc.date.available2022-03-07T16:33:45Z
dc.date.available2023-07-13T05:43:53Z
dc.date.issued2020-12
dc.identifier.urihttp://hdl.handle.net/20.500.14342/1003
dc.description.abstractHuman transthyretin (hTTR), a serum protein with a main role in transporting thyroid hormones and retinol through binding to the retinol-binding protein, is an amyloidogenic protein involved in familial amyloidotic polyneuropathy (FAP), familial amyloidotic cardiomyopathy, and central nervous system selective amyloidosis. hTTR also has a neuroprotective role in Alzheimer disease, being the major Aβ binding protein in human cerebrospinal fluid (CSF) that prevents amyloid-β (Aβ) aggregation with consequent abrogation of toxicity. Here we report an optimized preparative expression and purification protocol of hTTR (wt and amyloidogenic mutants) for in vitro screening assays of TTR ligands acting as amyloidogenesis inhibitors or acting as molecular chaperones to enhance the TTR:Aβ interaction. Preparative yields were up to 660 mg of homogenous protein per L of culture in fed-batch bioreactor. The recombinant wt protein is mainly unmodified at Cys10, the single cysteine in the protein sequence, whereas the highly amyloidogenic Y78F variant renders mainly the S-glutathionated form, which has essentially the same amyloidogenic behavior than the reduced protein with free Cys10. The TTR production protocol has shown inter-batch reproducibility of expression and protein quality for in vitro screening assays.eng
dc.format.extent12 p.cat
dc.language.isoengcat
dc.publisherMDPIcat
dc.relation.ispartofInternational Journal of Molecular Sciences. Vol.21, n.24 (2020), 9640cat
dc.rightsAttribution 4.0 International
dc.rights© L'autor/a
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/
dc.sourceRECERCAT (Dipòsit de la Recerca de Catalunya)
dc.subject.otherSèrumcat
dc.subject.otherHormones tiroidescat
dc.subject.otherRetinoidescat
dc.subject.otherProteïnescat
dc.subject.otherTransthyretincat
dc.subject.otherRecombinant expressioncat
dc.subject.otherFed-batch culturecat
dc.subject.otherProtein yieldcat
dc.subject.otherProtein-ligand interactionscat
dc.subject.otherProtein-protein interactionscat
dc.subject.otherAmyloid diseasescat
dc.titlePreparative scale production of recombinant human transthyretin for biophysical studies of protein-ligand and protein-protein interactionscat
dc.typeinfo:eu-repo/semantics/articlecat
dc.typeinfo:eu-repo/semantics/publishedVersioncat
dc.rights.accessLevelinfo:eu-repo/semantics/openAccess
dc.embargo.termscapcat
dc.subject.udc577
dc.identifier.doihttps://doi.org/10.3390/ijms21249640cat
dc.relation.projectIDinfo:eu-repo/grantAgreement/MINECO/PN I+D/PID2019-104350RB-I00cat
dc.relation.projectIDinfo:eu-repo/grantAgreement/SUR del DEC/SGR/2017-SGR-727cat
dc.relation.projectIDinfo:eu-repo/grantAgreement/Fundació La Marató TV3/20140330-31-32-33-34cat


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Except where otherwise noted, this item's license is described as http://creativecommons.org/licenses/by/4.0/
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