{ "dc.contributor.author": "Roura Padrosa, David" , "dc.contributor.author": "Castro Gallegos, Jessica" , "dc.contributor.author": "Romero Casañas, Alejandro" , "dc.contributor.author": "Ribó i Panosa, Marc" , "dc.contributor.author": "Vilanova i Brugués, Maria" , "dc.contributor.author": "Benito i Mundet, Antoni" , "dc.date.accessioned": "2019-01-10T10:24:38Z" , "dc.date.available": "2019-01-10T10:24:38Z" , "dc.date.issued": "2018-12-11" , "dc.identifier.issn": "1420-3049" , "dc.identifier.uri": "http://hdl.handle.net/10256/16185" , "dc.description.abstract": "Ribonucleases are proteins whose use is promising in anticancer therapy. We have previously constructed different human pancreatic ribonuclease variants that are selectively cytotoxic for tumor cells by introducing a nuclear localization signal into their sequence. However, these modifications produced an important decrease in their stability compromising their behavior in vivo. Here, we show that we can significantly increase the thermal stability of these cytotoxic proteins by introducing additional disulfide bonds by site-directed mutagenesis. One of these variants increases its thermal stability by around 17 °C, without affecting its catalytic activity while maintaining the cytotoxic activity against tumor cells. We also show that the most stable variant is significantly more resistant to proteolysis when incubated with proteinase K or with human sera, suggesting that its half-live could be increased in vivo once administered" , "dc.description.sponsorship": "This research was funded by MINECO (Spain), grant number BIO2013-43517 and by Universitat de Girona (Spain), grant number MPCU2016/18" , "dc.format.mimetype": "application/pdf" , "dc.language.iso": "eng" , "dc.publisher": "MDPI (Multidisciplinary Digital Publishing Institute)" , "dc.relation": "MINECO/PE 2014-2016/BIO2013-43517-R" , "dc.relation.isformatof": "Reproducció digital del document publicat a: https://doi.org/10.3390/molecules23123273" , "dc.relation.ispartof": "Molecules, 2018, vol. 23, núm. 12, p. 3273" , "dc.relation.ispartofseries": "Articles publicats (D-B)" , "dc.rights": "Attribution 4.0 International" , "dc.rights.uri": "http://creativecommons.org/licenses/by/4.0/" , "dc.subject": "Ribonucleases" , "dc.subject": "Citotoxicitat" , "dc.subject": "Cytotoxicity" , "dc.subject": "Càncer -- Tractament" , "dc.subject": "Cancer -- Treatment" , "dc.subject": "Cèl·lules canceroses" , "dc.subject": "Cancer cells" , "dc.title": "Construction of Highly Stable Cytotoxic Nuclear-Directed Ribonucleases" , "dc.type": "info:eu-repo/semantics/article" , "dc.rights.accessRights": "info:eu-repo/semantics/openAccess" , "dc.type.version": "info:eu-repo/semantics/publishedVersion" , "dc.identifier.doi": "https://doi.org/10.3390/molecules23123273" , "dc.contributor.funder": "Ministerio de Economía y Competitividad (Espanya)" , "dc.type.peerreviewed": "peer-reviewed" }