{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,14]],"date-time":"2026-09-14T09:06:13Z","timestamp":1789376773239,"version":"build-2803163510"},"reference-count":59,"publisher":"MDPI AG","issue":"9","license":[{"start":{"date-parts":[[2026,9,14]],"date-time":"2026-09-14T00:00:00Z","timestamp":1789344000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"the third batch of the \u201cTianchi Talent\u201d Young Doctoral Research Grant, Xinjiang Autonomous Region","award":["2025QNBS001"],"award-info":[{"award-number":["2025QNBS001"]}]},{"name":"the Talent Team Support Initiative for Scientific Research and Innovation Platforms of the Xinjiang Talent Development Fund","award":["XJRCXYCTD2025-2"],"award-info":[{"award-number":["XJRCXYCTD2025-2"]}]},{"award":["XJRCXYCTD2025-2"],"award-info":[{"award-number":["XJRCXYCTD2025-2"]}],"id":[{"id":"https:\/\/ror.org\/01rkhv978","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biomolecules"],"abstract":"<jats:p>Tuberculosis (TB), caused by Mycobacterium tuberculosis (Mtb), remains a major global health concern, particularly due to the emergence of drug-resistant strains. FAD-containing monooxygenase EthA activates the antitubercular prodrug ethionamide (ETH) in Mtb. However, the structural and functional mechanisms of Mtb EthA are not fully understood. Here, we report an AlphaFold2-predicted structural model of Mtb EthA, validated using Ramachandran analysis (91.7% residues in favored regions) and a ProSA Z-score of \u221210.96. Based on the results of molecular docking, site-directed mutagenesis was conducted. We found that R207 is required for EthA activity. Alanine substitutions at T186, S208, and T210 resulted in 5.8- to 7.2-fold reductions in activity. Conversely, deletion of three peripheral segments (residues 137\u2013176, 315\u2013336, and 419\u2013460) enhanced activity by 1.9-, 1.5-, and 1.2-fold, respectively. These results provide a structural framework for EthA function and suggest that flexible peripheral regions may constrain catalytic activity, offering insights into ETH activation and potential mechanisms of drug resistance.<\/jats:p>","DOI":"10.3390\/biom16091333","type":"journal-article","created":{"date-parts":[[2026,9,14]],"date-time":"2026-09-14T08:10:23Z","timestamp":1789373423000},"page":"1333","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["The Flexible Peripheral Segments Modulate the Catalytic Activity of the FAD-Containing Monooxygenase EthA from Mycobacterium tuberculosis"],"prefix":"10.3390","volume":"16","author":[{"given":"Dafeng","family":"Liu","sequence":"first","affiliation":[{"name":"College of Biological Sciences and Technology, Yili Normal University, Yining 835000, China"},{"name":"School of Life Sciences, Xiamen University, Xiamen 361102, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"conceptualization","vocabulary":"credit"},{"role":"investigation","vocabulary":"credit"},{"role":"writing-review-editing","vocabulary":"credit"},{"role":"funding-acquisition","vocabulary":"credit"},{"role":"visualization","vocabulary":"credit"},{"role":"resources","vocabulary":"credit"},{"role":"methodology","vocabulary":"credit"},{"role":"software","vocabulary":"credit"},{"role":"data-curation","vocabulary":"credit"},{"role":"supervision","vocabulary":"credit"},{"role":"project-administration","vocabulary":"credit"},{"role":"writing-original-draft","vocabulary":"credit"},{"role":"validation","vocabulary":"credit"},{"role":"formal-analysis","vocabulary":"credit"}]},{"given":"Xieping","family":"Sun","sequence":"additional","affiliation":[{"name":"College of Biological Sciences and Technology, Yili Normal University, Yining 835000, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"investigation","vocabulary":"credit"}]},{"given":"Feng","family":"Yu","sequence":"additional","affiliation":[{"name":"College of Biological Sciences and Technology, Yili Normal University, Yining 835000, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"investigation","vocabulary":"credit"}]},{"given":"Hongying","family":"Song","sequence":"additional","affiliation":[{"name":"School of Life Sciences, Xiamen University, Xiamen 361102, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"investigation","vocabulary":"credit"}]},{"given":"Wenshuang","family":"Yao","sequence":"additional","affiliation":[{"name":"School of Life Sciences, Xiamen University, Xiamen 361102, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"investigation","vocabulary":"credit"}]},{"given":"Huashui","family":"Deng","sequence":"additional","affiliation":[{"name":"School of Life Sciences, Xiamen University, Xiamen 361102, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"investigation","vocabulary":"credit"}]},{"given":"Daoqi","family":"Song","sequence":"additional","affiliation":[{"name":"School of Life Sciences, Xiamen University, Xiamen 361102, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"investigation","vocabulary":"credit"}]}],"member":"1968","published-online":{"date-parts":[[2026,9,14]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"850","DOI":"10.1016\/S0140-6736(24)02479-6","article-title":"Tuberculosis","volume":"405","author":"Trajman","year":"2025","journal-title":"Lancet"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1789","DOI":"10.1056\/NEJMoa2412381","article-title":"BCG Revaccination for the Prevention of Mycobacterium tuberculosis Infection","volume":"392","author":"Schmidt","year":"2025","journal-title":"N. 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