{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,26]],"date-time":"2026-08-26T06:25:09Z","timestamp":1787725509414,"version":"build-2784847793"},"reference-count":51,"publisher":"Wiley","license":[{"start":{"date-parts":[[2026,8,25]],"date-time":"2026-08-25T00:00:00Z","timestamp":1787616000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"},{"start":{"date-parts":[[2026,8,25]],"date-time":"2026-08-25T00:00:00Z","timestamp":1787616000000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"funder":[{"DOI":"10.13039\/100010665","name":"H2020 Marie Sk\u0142odowska-Curie Actions","doi-asserted-by":"publisher","award":["847675 COFUND project oLife"],"award-info":[{"award-number":["847675 COFUND project oLife"]}],"id":[{"id":"10.13039\/100010665","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100002923","name":"Consejo Nacional de Investigaciones Cient\u00edficas y T\u00e9cnicas","doi-asserted-by":"publisher","award":["PIP\u20100409CO"],"award-info":[{"award-number":["PIP\u20100409CO"]}],"id":[{"id":"10.13039\/501100002923","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100002923","name":"Consejo Nacional de Investigaciones Cient\u00edficas y T\u00e9cnicas","doi-asserted-by":"publisher","award":["Reallocation allowance RESOL\u20102024\u20101665\u2010APN\u2010DIR#CO"],"award-info":[{"award-number":["Reallocation allowance RESOL\u20102024\u20101665\u2010APN\u2010DIR#CO"]}],"id":[{"id":"10.13039\/501100002923","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["febs.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["The FEBS Journal"],"abstract":"<jats:p>\n                    Type II NADH dehydrogenases (NDH\u20102) are accessory enzymes of the bacterial electron transport chain (ETC). While they functionally overlap with complex I, their main role is not proton translocation but maintaining the intracellular NADH\/NAD\n                    <jats:sup>+<\/jats:sup>\n                    balance. Although often non\u2010essential, NDH\u20102 become crucial in species lacking complex I, serving as the primary electron entry point into the ETC. Their virtual absence in mammals makes these enzymes attractive targets for antimicrobial drug development and mitochondrial functional restoration. NDH\u20102 catalyse electron transfer from NADH to quinones, yet two distinct catalytic mechanisms have been described for members of the family: a classical ping\u2010pong one or an atypical ternary mechanism involving the formation of a charge transfer complex (CTC). The molecular basis of these mechanisms remains unclear. Additionally, their occurrence among NDH\u20102 from different bacterial lineages is unknown. Here we combined molecular phylogenetics, ancestral sequence reconstruction, expression and biochemical characterisation of ancestral and modern enzymes, and molecular dynamics simulations to explore the mechanistic versatility of NDH\u20102 across Bacteria. Our results show the atypical ternary mechanism is restricted to the Firmicutes (Bacillota) lineage and correlates with the presence of a single substitution located at the bottom of the active site. This work provides an evolutionary framework for understanding the mechanistic versatility of NDH\u20102. Furthermore, it establishes a basis for drug discovery targeting pathogenic strains and opens avenues for developing innovative strategies to complement dysfunctional mitochondria.\n                  <\/jats:p>","DOI":"10.1111\/febs.70697","type":"journal-article","created":{"date-parts":[[2026,8,26]],"date-time":"2026-08-26T05:40:57Z","timestamp":1787722857000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["The evolution and mechanistic versatility of the bacterial\n                    <scp>NADH<\/scp>\n                    dehydrogenases type\n                    <scp>II<\/scp>"],"prefix":"10.1111","author":[{"given":"Diego","family":"Masone","sequence":"first","affiliation":[{"name":"IHEM, CONICET\u2010UNCuyo  Mendoza Argentina"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lianne","family":"van Es","sequence":"additional","affiliation":[{"name":"Molecular Enzymology group University of Groningen  The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Guang","family":"Yang","sequence":"additional","affiliation":[{"name":"Molecular Enzymology group University of Groningen  The Netherlands"},{"name":"School of Food Science and Pharmaceutical Engineering Nanjing Normal University  China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"C. Marcelo","family":"Roggero","sequence":"additional","affiliation":[{"name":"IHEM, CONICET\u2010UNCuyo  Mendoza Argentina"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6346-5014","authenticated-orcid":false,"given":"Marco W.","family":"Fraaije","sequence":"additional","affiliation":[{"name":"Molecular Enzymology group University of Groningen  The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5817-4631","authenticated-orcid":false,"given":"Mar\u00eda Laura","family":"Mascotti","sequence":"additional","affiliation":[{"name":"IHEM, CONICET\u2010UNCuyo  Mendoza Argentina"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,8,25]]},"reference":[{"key":"e_1_2_10_2_1","doi-asserted-by":"crossref","first-page":"331","DOI":"10.1016\/bs.ampbs.2019.03.002","article-title":"The plethora of membrane respiratory chains in the phyla 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