{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,16]],"date-time":"2026-07-16T04:42:54Z","timestamp":1784176974678,"version":"3.55.0"},"reference-count":37,"publisher":"Public Library of Science (PLoS)","issue":"10","license":[{"start":{"date-parts":[[2021,10,13]],"date-time":"2021-10-13T00:00:00Z","timestamp":1634083200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000062","name":"National Institute of Diabetes and Digestive and Kidney Diseases","doi-asserted-by":"publisher","award":["DK090242"],"award-info":[{"award-number":["DK090242"]}],"id":[{"id":"10.13039\/100000062","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS ONE"],"abstract":"<jats:p>Mitochondrial trifunctional protein (TFP) is a membrane-associated heterotetramer that catalyzes three of the four reactions needed to chain-shorten long-chain fatty acids inside the mitochondria. TFP is known to be heavily modified by acetyllysine and succinyllysine post-translational modifications (PTMs), many of which are targeted for reversal by the mitochondrial sirtuin deacylases SIRT3 and SIRT5. However, the functional significance of these PTMs is not clear, with some reports showing TFP gain-of-function and some showing loss-of-function upon increased acylation. Here, we mapped the known SIRT3\/SIRT5-targeted lysine residues onto the recently solved TFP crystal structure which revealed that many of the target sites are involved in substrate channeling within the TFP\u03b1 subunit. To test the effects of acylation on substate channeling through TFP\u03b1, we enzymatically synthesized the physiological long-chain substrate (2E)-hexadecenoyl-CoA. Assaying TFP in SIRT3 and SIRT5 knockout mouse liver and heart mitochondria with (2E)-hexadecenoyl-CoA revealed no change in enzyme activity. Finally, we investigated the effects of lysine acylation on TFP membrane binding <jats:italic>in vitro<\/jats:italic>. Acylation did not alter recombinant TFP binding to cardiolipin-containing liposomes. However, the presence of liposomes strongly abrogated the acylation reaction between succinyl-CoA and TFP lysine residues. Thus, TFP in the membrane-bound state may be protected against lysine acylation.<\/jats:p>","DOI":"10.1371\/journal.pone.0256619","type":"journal-article","created":{"date-parts":[[2021,10,13]],"date-time":"2021-10-13T18:45:17Z","timestamp":1634150717000},"page":"e0256619","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":8,"title":["The enzyme activity of mitochondrial trifunctional protein is not altered by lysine acetylation or lysine succinylation"],"prefix":"10.1371","volume":"16","author":[{"given":"Yuxun","family":"Zhang","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9476-309X","authenticated-orcid":true,"given":"Eric","family":"Goetzman","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2021,10,13]]},"reference":[{"key":"pone.0256619.ref001","doi-asserted-by":"crossref","first-page":"132","DOI":"10.1007\/s40142-017-0125-6","article-title":"Advances in the Understanding and Treatment of Mitochondrial Fatty Acid Oxidation Disorders","volume":"5","author":"ES Goetzman","year":"2017","journal-title":"Curr Genet Med Rep"},{"key":"pone.0256619.ref002","doi-asserted-by":"crossref","first-page":"317","DOI":"10.1016\/j.pcl.2017.11.006","article-title":"Inborn Errors of Metabolism with Myopathy: Defects of Fatty Acid Oxidation and the Carnitine 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