dc.contributor.author | Carrasco del Amor, Ana María | |
dc.contributor.author | Freitas, Sara | |
dc.contributor.author | Urbatzka, Ralph | |
dc.contributor.author | Fresnedo Aranguren, María Olatz | |
dc.contributor.author | Cristobal Barragán, Susana | |
dc.date.accessioned | 2020-02-07T09:29:52Z | |
dc.date.available | 2020-02-07T09:29:52Z | |
dc.date.issued | 2019-06-21 | |
dc.identifier.citation | Marine Drugs 17(6) : (2019) // Article ID 371 | es_ES |
dc.identifier.issn | 1660-3397 | |
dc.identifier.uri | http://hdl.handle.net/10810/40506 | |
dc.description.abstract | The acceleration of the process of understanding the pharmacological application of new marine bioactive compounds requires identifying the compound protein targets leading the molecular mechanisms in a living cell. The thermal proteome profiling (TPP) methodology does not fulfill the requirements for its application to any bioactive compound lacking chemical and functional characterization. Here, we present a modified method that we called bTPP for bioactive thermal proteome profiling that guarantees target specificity from a soluble subproteome. We showed that the precipitation of the microsomal fraction before the thermal shift assay is crucial to accurately calculate the melting points of the protein targets. As a probe of concept, the protein targets of 13(2)-hydroxy-pheophytin, a compound previously isolated from a marine cyanobacteria for its lipid reducing activity, were analyzed on the hepatic cell line HepG2. Our improved method identified 9 protein targets out of 2500 proteins, including 3 targets (isocitrate dehydrogenase, aldehyde dehydrogenase, phosphoserine aminotransferase) that could be related to obesity and diabetes, as they are involved in the regulation of insulin sensitivity and energy metabolism. This study demonstrated that the bTPP method can accelerate the field of biodiscovery, revealing protein targets involved in mechanisms of action (MOA) connected with future applications of bioactive compounds. | es_ES |
dc.description.sponsorship | This project received funding from the ERA-NET Marine Biotechnology project CYANOBESITY that it is cofounding from FORMAS, Sweden grant nr. 2016-02004 (SC), FCT Foundation of Science and Technology, Portugal, grant number ERA-MBT/0001/2015 (RU). This work has also been funded by IKERBASQUE (SC), Basque Government grant IT-971-16 (SC and OF), and FCT grants SFRH/BPD/112287/2015, SFRH/BD/116009/2016, FCT strategic fund (UID/Multi/04423/2019) (RU and SF). | es_ES |
dc.language.iso | eng | es_ES |
dc.publisher | MDPI | es_ES |
dc.rights | info:eu-repo/semantics/openAccess | es_ES |
dc.rights.uri | http://creativecommons.org/licenses/by/3.0/es/ | * |
dc.subject | thermal proteome profiling | es_ES |
dc.subject | mechanisms of action | es_ES |
dc.subject | bioactive compound | es_ES |
dc.subject | label-free quantitative proteomics | es_ES |
dc.subject | marine biodiscovery | es_ES |
dc.subject | insulin sensitivity | es_ES |
dc.subject | target | es_ES |
dc.subject | purification | es_ES |
dc.subject | cells | es_ES |
dc.subject | mice | es_ES |
dc.title | Application of Bioactive Thermal Proteome Profiling to Decipher the Mechanism of Action of the Lipid Lowering 13(2)-Hydroxy-pheophytin Isolated from a Marine Cyanobacteria | es_ES |
dc.type | info:eu-repo/semantics/article | es_ES |
dc.rights.holder | This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0) | es_ES |
dc.rights.holder | Atribución 3.0 España | * |
dc.relation.publisherversion | https://www.mdpi.com/1660-3397/17/6/371 | es_ES |
dc.identifier.doi | 10.3390/md17060371 | |
dc.departamentoes | Fisiología | es_ES |
dc.departamentoeu | Fisiologia | es_ES |