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http://hdl.handle.net/10261/225741
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Campo DC | Valor | Lengua/Idioma |
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dc.contributor.author | Cotrina, Ellen Y. | - |
dc.contributor.author | Vilà, Marta | - |
dc.contributor.author | Nieto, Joan | - |
dc.contributor.author | Arsequell, Gemma | - |
dc.contributor.author | Planas, Antoni | - |
dc.date.accessioned | 2020-12-28T14:28:01Z | - |
dc.date.available | 2020-12-28T14:28:01Z | - |
dc.date.issued | 2020 | - |
dc.identifier.citation | International Journal of Molecular Sciences 21(24): 9640 (2020) | - |
dc.identifier.uri | http://hdl.handle.net/10261/225741 | - |
dc.description | © 2020 by the authors. | - |
dc.description.abstract | Human 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. | - |
dc.description.sponsorship | Work supported in part by grant PID2019-104350RB-I00 from MINECO and grant 2017SGR-727 from the Generalitat de Catalunya (to A.P.). G.A. acknowledges a grant from Fundació Marató de TV3, Spain (Project ref. 20140330-31-32-33-34). E.Y.C. acknowledges a contract funded by the project of Fundació Marató de TV3 and a contract from Ford España—Fundación Apadrina la Ciencia. | - |
dc.language.iso | eng | - |
dc.publisher | Multidisciplinary Digital Publishing Institute | - |
dc.relation.isversionof | Publisher's version | - |
dc.rights | openAccess | - |
dc.subject | Transthyretin | - |
dc.subject | Recombinant expression | - |
dc.subject | Fed-batch culture | - |
dc.subject | Protein yield | - |
dc.subject | Protein-ligand interactions | - |
dc.subject | Protein-protein interactions | - |
dc.subject | Amyloid diseases | - |
dc.subject | Generalitat de Catalunya | - |
dc.subject | Fundació La Marató de TV3 | - |
dc.subject | Ford España | - |
dc.title | Preparative Scale Production of Recombinant Human Transthyretin for Biophysical Studies of Protein-Ligand and Protein-Protein Interactions | - |
dc.type | artículo | - |
dc.identifier.doi | 10.3390/ijms21249640 | - |
dc.description.peerreviewed | Peer reviewed | - |
dc.relation.publisherversion | https://doi.org/10.3390/ijms21249640 | - |
dc.identifier.e-issn | 1422-0067 | - |
dc.date.updated | 2020-12-28T14:28:02Z | - |
dc.rights.license | http://creativecommons.org/licenses/by/4.0/ | - |
dc.relation.csic | Sí | - |
dc.identifier.pmid | 33348885 | - |
dc.type.coar | http://purl.org/coar/resource_type/c_6501 | es_ES |
item.fulltext | With Fulltext | - |
item.languageiso639-1 | en | - |
item.openairecristype | http://purl.org/coar/resource_type/c_18cf | - |
item.openairetype | artículo | - |
item.cerifentitytype | Publications | - |
item.grantfulltext | open | - |
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Preparative_Cotrina_Art2020.pdf | 1,87 MB | Adobe PDF | Visualizar/Abrir |
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