{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,14]],"date-time":"2026-07-14T12:02:24Z","timestamp":1784030544921,"version":"3.55.0"},"reference-count":93,"publisher":"MDPI AG","issue":"4","license":[{"start":{"date-parts":[[2019,2,16]],"date-time":"2019-02-16T00:00:00Z","timestamp":1550275200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001871","name":"Funda\u00e7\u00e3o para a Ci\u00eancia e a Tecnologia","doi-asserted-by":"publisher","award":["UID\/Multi\/04378\/2013; SFRH\/BPD\/97719\/2013"],"award-info":[{"award-number":["UID\/Multi\/04378\/2013; SFRH\/BPD\/97719\/2013"]}],"id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]},{"name":"ERDF under the PT2020 Partnership Agreement","award":["POCI-01-0145-FEDER-007728"],"award-info":[{"award-number":["POCI-01-0145-FEDER-007728"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Bcl-2 protein is involved in cell apoptosis and is considered an interesting target for anti-cancer therapy. The present study aims to understand the stability and conformational changes of Bcl-2 upon interaction with the inhibitor venetoclax, and to explore other drug-target regions. We combined biophysical and in silico approaches to understand the mechanism of ligand binding to Bcl-2. Thermal shift assay (TSA) and urea electrophoresis showed a significant increase in protein stability upon venetoclax incubation, which is corroborated by molecular docking and molecular dynamics simulations. An 18 \u00b0C shift in Bcl-2 melting temperature was observed in the TSA, corresponding to a binding affinity multiple times higher than that of any other reported Bcl-2 inhibitor. This protein-ligand interaction does not implicate alternations in protein conformation, as suggested by SAXS. Additionally, bioinformatics approaches were used to identify deleterious non-synonymous single nucleotide polymorphisms (nsSNPs) of Bcl-2 and their impact on venetoclax binding, suggesting that venetoclax interaction is generally favored against these deleterious nsSNPs. Apart from the BH3 binding groove of Bcl-2, the flexible loop domain (FLD) also plays an important role in regulating the apoptotic process. High-throughput virtual screening (HTVS) identified 5 putative FLD inhibitors from the Zinc database, showing nanomolar affinity toward the FLD of Bcl-2.<\/jats:p>","DOI":"10.3390\/ijms20040860","type":"journal-article","created":{"date-parts":[[2019,2,17]],"date-time":"2019-02-17T22:11:50Z","timestamp":1550441510000},"page":"860","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":32,"title":["Shedding Light on the Interaction of Human Anti-Apoptotic Bcl-2 Protein with Ligands through Biophysical and in Silico Studies"],"prefix":"10.3390","volume":"20","author":[{"given":"Joao","family":"Ramos","sequence":"first","affiliation":[{"name":"UCIBIO-NOVA, Departamento de Qu\u00edmica, Faculdade de Ci\u00eancias e Tecnologia, Universidade NOVA de Lisboa, 2829-516 Caparica, Portugal"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0953-1196","authenticated-orcid":false,"given":"Jayaraman","family":"Muthukumaran","sequence":"additional","affiliation":[{"name":"UCIBIO-NOVA, Departamento de Qu\u00edmica, Faculdade de Ci\u00eancias e Tecnologia, Universidade NOVA de Lisboa, 2829-516 Caparica, Portugal"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5310-0668","authenticated-orcid":false,"given":"Filipe","family":"Freire","sequence":"additional","affiliation":[{"name":"UCIBIO-NOVA, Departamento de Qu\u00edmica, Faculdade de Ci\u00eancias e Tecnologia, Universidade NOVA de Lisboa, 2829-516 Caparica, Portugal"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jo\u00e3o","family":"Paquete-Ferreira","sequence":"additional","affiliation":[{"name":"UCIBIO-NOVA, Departamento de Qu\u00edmica, Faculdade de Ci\u00eancias e Tecnologia, Universidade NOVA de Lisboa, 2829-516 Caparica, Portugal"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ana Rita","family":"Otrelo-Cardoso","sequence":"additional","affiliation":[{"name":"UCIBIO-NOVA, Departamento de Qu\u00edmica, Faculdade de Ci\u00eancias e Tecnologia, Universidade NOVA de Lisboa, 2829-516 Caparica, Portugal"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dmitri","family":"Svergun","sequence":"additional","affiliation":[{"name":"European Molecular Biology Laboratory (EMBL), Hamburg Outstation, c\/o DESY, 22067 Hamburg, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alejandro","family":"Panjkovich","sequence":"additional","affiliation":[{"name":"European Molecular Biology Laboratory (EMBL), Hamburg Outstation, c\/o DESY, 22067 Hamburg, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Teresa","family":"Santos-Silva","sequence":"additional","affiliation":[{"name":"UCIBIO-NOVA, Departamento de Qu\u00edmica, Faculdade de Ci\u00eancias e Tecnologia, Universidade NOVA de Lisboa, 2829-516 Caparica, Portugal"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2019,2,16]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"57","DOI":"10.1016\/S0092-8674(00)81683-9","article-title":"The Hallmarks of Cancer","volume":"100","author":"Hanahan","year":"2000","journal-title":"Cell"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"646","DOI":"10.1016\/j.cell.2011.02.013","article-title":"Hallmarks of cancer: The next generation","volume":"144","author":"Hanahan","year":"2011","journal-title":"Cell"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1214","DOI":"10.1111\/j.1582-4934.2007.00150.x","article-title":"Cell death in health and disease","volume":"11","author":"Lockshin","year":"2007","journal-title":"J. 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