{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,8]],"date-time":"2026-07-08T11:40:46Z","timestamp":1783510846280,"version":"3.55.0"},"reference-count":31,"publisher":"MDPI AG","issue":"1","license":[{"start":{"date-parts":[[2024,1,11]],"date-time":"2024-01-11T00:00:00Z","timestamp":1704931200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100007065","name":"University of Salerno","doi-asserted-by":"publisher","award":["ORSA215145"],"award-info":[{"award-number":["ORSA215145"]}],"id":[{"id":"10.13039\/501100007065","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100007065","name":"University of Salerno","doi-asserted-by":"publisher","award":["2022RCTY7M"],"award-info":[{"award-number":["2022RCTY7M"]}],"id":[{"id":"10.13039\/501100007065","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100007065","name":"University of Salerno","doi-asserted-by":"publisher","award":["P2022CKMPW"],"award-info":[{"award-number":["P2022CKMPW"]}],"id":[{"id":"10.13039\/501100007065","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Ministero dell\u2019Universit\u00e0 e della Ricerca (MUR)","award":["ORSA215145"],"award-info":[{"award-number":["ORSA215145"]}]},{"name":"Ministero dell\u2019Universit\u00e0 e della Ricerca (MUR)","award":["2022RCTY7M"],"award-info":[{"award-number":["2022RCTY7M"]}]},{"name":"Ministero dell\u2019Universit\u00e0 e della Ricerca (MUR)","award":["P2022CKMPW"],"award-info":[{"award-number":["P2022CKMPW"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Marine Drugs"],"abstract":"<jats:p>Mass spectrometry-based chemical proteomic approaches using limited proteolysis have become a powerful tool for the identification and analysis of the interactions between a small molecule (SM) and its protein target(s). Gracilioether A (GeA) is a polyketide isolated from a marine sponge, for which we aimed to trace the interactome using this strategy. DARTS (Drug Affinity Responsive Target Stability) and t-LiP-MS (targeted-Limited Proteolysis-Mass Spectrometry) represented the main techniques used in this study. DARTS was applied on HeLa cell lysate for the identification of the GeA target proteins, and t-LiP-MS was employed to investigate the protein\u2019s regions involved in the binding with GeA. The results were complemented through the use of binding studies using Surface Plasmon Resonance (SPR) and in silico molecular docking experiments. Ubiquitin carboxyl-terminal hydrolase 5 (USP5) was identified as a promising target of GeA, and the interaction profile of the USP5-GeA complex was explained. USP5 is an enzyme involved in the pathway of protein metabolism through the disassembly of the polyubiquitin chains on degraded proteins into ubiquitin monomers. This activity is connected to different cellular functions concerning the maintenance of chromatin structure and receptors and the degradation of abnormal proteins and cancerogenic progression. On this basis, this structural information opens the way to following studies focused on the definition of the biological potential of Gracilioether A and the rational development of novel USP5 inhibitors based on a new structural skeleton.<\/jats:p>","DOI":"10.3390\/md22010041","type":"journal-article","created":{"date-parts":[[2024,1,11]],"date-time":"2024-01-11T12:56:33Z","timestamp":1704977793000},"page":"41","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":14,"title":["Chemoproteomics Reveals USP5 (Ubiquitin Carboxyl-Terminal Hydrolase 5) as Promising Target of the Marine Polyketide Gracilioether A"],"prefix":"10.3390","volume":"22","author":[{"given":"Alessandra","family":"Capuano","sequence":"first","affiliation":[{"name":"Dipartimento di Farmacia, University of Salerno, Via Giovanni Paolo II 132, 84084 Fisciano, Salerno, Italy"},{"name":"PhD Program in Drug Discovery and Development, University of Salerno, 84084 Fisciano, Salerno, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4407-2781","authenticated-orcid":false,"given":"Gilda","family":"D\u2019Urso","sequence":"additional","affiliation":[{"name":"Dipartimento di Farmacia, University of Salerno, Via Giovanni Paolo II 132, 84084 Fisciano, Salerno, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michela","family":"Aliberti","sequence":"additional","affiliation":[{"name":"Dipartimento di Farmacia, University of Salerno, Via Giovanni Paolo II 132, 84084 Fisciano, Salerno, Italy"},{"name":"PhD Program in Drug Discovery and Development, University of Salerno, 84084 Fisciano, Salerno, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dafne","family":"Ruggiero","sequence":"additional","affiliation":[{"name":"Dipartimento di Farmacia, University of Salerno, Via Giovanni Paolo II 132, 84084 Fisciano, Salerno, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Stefania","family":"Terracciano","sequence":"additional","affiliation":[{"name":"Dipartimento di Farmacia, University of Salerno, Via Giovanni Paolo II 132, 84084 Fisciano, Salerno, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8879-4332","authenticated-orcid":false,"given":"Carmen","family":"Festa","sequence":"additional","affiliation":[{"name":"Dipartimento di Farmacia, University of Napoli \u201cFederico II\u201d, Via Domenico Montesano 49, 80131 Napoli, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5443-1565","authenticated-orcid":false,"given":"Alessandra","family":"Tosco","sequence":"additional","affiliation":[{"name":"Dipartimento di Farmacia, University of Salerno, Via Giovanni Paolo II 132, 84084 Fisciano, Salerno, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maria Giovanna","family":"Chini","sequence":"additional","affiliation":[{"name":"Dipartimento di Bioscienze e Territorio, University of Molise, Contrada Fonte Lappone, 86090 Isernia, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5065-9717","authenticated-orcid":false,"given":"Gianluigi","family":"Lauro","sequence":"additional","affiliation":[{"name":"Dipartimento di Farmacia, University of Salerno, Via Giovanni Paolo II 132, 84084 Fisciano, Salerno, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Giuseppe","family":"Bifulco","sequence":"additional","affiliation":[{"name":"Dipartimento di Farmacia, University of Salerno, Via Giovanni Paolo II 132, 84084 Fisciano, Salerno, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2310-9426","authenticated-orcid":false,"given":"Agostino","family":"Casapullo","sequence":"additional","affiliation":[{"name":"Dipartimento di Farmacia, University of Salerno, Via Giovanni Paolo II 132, 84084 Fisciano, Salerno, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2024,1,11]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1937","DOI":"10.1021\/jacs.7b12903","article-title":"Enantioselective Total Syntheses of (+)-Hippolachnin A, (+)-Gracilioether A, (\u2212)-Gracilioether E, and (\u2212)-Gracilioether F","volume":"140","author":"Li","year":"2018","journal-title":"J. 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