120016 20240319081002.0 doi 10.1002/pro.4427 sideral 130894 ART-2022-130894 eng Jimenez-Alesanco, Ana Universidad de Zaragoza (orcid)0000-0003-4726-7821 Repositioning small molecule drugs as allosteric inhibitors of the BFT-3 toxin from enterotoxigenic Bacteroides fragilis 2022 Access copy available to the general public Unrestricted Bacteroides fragilis is an abundant commensal component of the healthy human colon. However, under dysbiotic conditions, enterotoxigenic B. fragilis (ETBF) may arise and elicit diarrhea, anaerobic bacteremia, inflammatory bowel disease, and colorectal cancer. Most worrisome, ETBF is resistant to many disparate antibiotics. ETBF's only recognized specific virulence factor is a zinc-dependent metallopeptidase (MP) called B. fragilis toxin (BFT) or fragilysin, which damages the intestinal mucosa and triggers disease-related signaling mechanisms. Thus, therapeutic targeting of BFT is expected to limit ETBF pathogenicity and improve the prognosis for patients. We focused on one of the naturally occurring BFT isoforms, BFT-3, and managed to repurpose several approved drugs as BFT-3 inhibitors through a combination of biophysical, biochemical, structural, and cellular techniques. In contrast to canonical MP inhibitors, which target the active site of mature enzymes, these effectors bind to a distal allosteric site in the proBFT-3 zymogen structure, which stabilizes a partially unstructured, zinc-free enzyme conformation by shifting a zinc-dependent disorder-to-order equilibrium. This yields proBTF-3 incompetent for autoactivation, thus ablating hydrolytic activity of the mature toxin. Additionally, a similar destabilizing effect is observed for the activated protease according to biophysical and biochemical data. Our strategy paves a novel way for the development of highly specific inhibitors of ETBF-mediated enteropathogenic conditions. info:eu-repo/grantAgreement/ES/DGA/E25-20R info:eu-repo/semantics/openAccess by-nc-nd http://creativecommons.org/licenses/by-nc-nd/3.0/es/ 8.0 2022 BIOCHEMISTRY & MOLECULAR BIOLOGY 36 / 285 = 0.126 2022 Q1 T1 4.007 2022 Biochemistry 2022 Q1 Molecular Biology 2022 Q1 Medicine (miscellaneous) 2022 Q1 10.8 2022 info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion Eckhard, Ulrich Asencio del Rio, Marta Vega, Sonia (orcid)0000-0002-1232-6310 Guevara, Tibisay Velazquez-Campoy, Adrian Universidad de Zaragoza (orcid)0000-0001-5702-4538 Gomis-Rüth, Francesc Xavier Abian, Olga Universidad de Zaragoza (orcid)0000-0001-5664-1729 1002 060 Universidad de Zaragoza Dpto. Bioq.Biolog.Mol. Celular Área Bioquímica y Biolog.Mole. 31, 10 (2022), [18 pp.] Protein sci. Protein science 0961-8368 4884677 http://zaguan.unizar.es/record/120016/files/texto_completo.pdf Versión publicada 2642195 http://zaguan.unizar.es/record/120016/files/texto_completo.jpg?subformat=icon icon Versión publicada oai:zaguan.unizar.es:120016 articulos driver 2024-03-18-14:16:16 ARTICLE