{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,16]],"date-time":"2026-04-16T22:22:44Z","timestamp":1776378164725,"version":"3.51.2"},"reference-count":31,"publisher":"MDPI AG","issue":"6","license":[{"start":{"date-parts":[[2019,6,21]],"date-time":"2019-06-21T00:00:00Z","timestamp":1561075200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001862","name":"Svenska Forskningsr\u00e5det Formas","doi-asserted-by":"publisher","award":["2016-02004"],"award-info":[{"award-number":["2016-02004"]}],"id":[{"id":"10.13039\/501100001862","id-type":"DOI","asserted-by":"publisher"}]},{"name":"FCT grants SFRH\/BPD","award":["112287\/2015"],"award-info":[{"award-number":["112287\/2015"]}]},{"name":"FCT Foundation of Science and Technology, Portugal","award":["ERA-MBT\/0001\/2015"],"award-info":[{"award-number":["ERA-MBT\/0001\/2015"]}]},{"name":"Basque Government grant","award":["IT-971-16"],"award-info":[{"award-number":["IT-971-16"]}]},{"name":"SFRH\/BD","award":["SFRH\/BD\/116009\/2016"],"award-info":[{"award-number":["SFRH\/BD\/116009\/2016"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Marine Drugs"],"abstract":"<jats:p>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 132-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.<\/jats:p>","DOI":"10.3390\/md17060371","type":"journal-article","created":{"date-parts":[[2019,6,21]],"date-time":"2019-06-21T11:54:31Z","timestamp":1561118071000},"page":"371","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":19,"title":["Application of Bioactive Thermal Proteome Profiling to Decipher the Mechanism of Action of the Lipid Lowering 132-Hydroxy-pheophytin Isolated from a Marine Cyanobacteria"],"prefix":"10.3390","volume":"17","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-5145-9327","authenticated-orcid":false,"given":"Ana","family":"Carrasco del Amor","sequence":"first","affiliation":[{"name":"Department of Clinical and Experimental Medicine, Cell Biology, Faculty of Medicine, Link\u00f6ping University, 581 85 Link\u00f6ping, Sweden"}]},{"given":"Sara","family":"Freitas","sequence":"additional","affiliation":[{"name":"CIIMAR, Interdisciplinary Centre of Marine and Environmental Research, 4450-208 Matosinhos, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7476-9195","authenticated-orcid":false,"given":"Ralph","family":"Urbatzka","sequence":"additional","affiliation":[{"name":"CIIMAR, Interdisciplinary Centre of Marine and Environmental Research, 4450-208 Matosinhos, Portugal"}]},{"given":"Olatz","family":"Fresnedo","sequence":"additional","affiliation":[{"name":"Department of Physiology, Faculty of Medicine and Nursing, University of the Basque Country UPV\/EHU, 48940 Leioa, Spain"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3894-2218","authenticated-orcid":false,"given":"Susana","family":"Cristobal","sequence":"additional","affiliation":[{"name":"Department of Clinical and Experimental Medicine, Cell Biology, Faculty of Medicine, Link\u00f6ping University, 581 85 Link\u00f6ping, Sweden"},{"name":"Department of Physiology, Ikerbasque, Faculty of Medicine and Nursing, University of the Basque Country UPV\/EH, 48940 Leioa, Spain"}]}],"member":"1968","published-online":{"date-parts":[[2019,6,21]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"531","DOI":"10.1038\/nrd.2017.111","article-title":"Opportunities and challenges in phenotypic drug discovery: An industry perspective","volume":"16","author":"Moffat","year":"2017","journal-title":"Nat. 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