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The enzyme activity of mitochondrial trifunctional protein is not altered by lysine acetylation or lysine succinylation

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...

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Detalles Bibliográficos
Autores principales: Zhang, Yuxun, Goetzman, Eric
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8513871/
https://www.ncbi.nlm.nih.gov/pubmed/34644302
http://dx.doi.org/10.1371/journal.pone.0256619
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author Zhang, Yuxun
Goetzman, Eric
author_facet Zhang, Yuxun
Goetzman, Eric
author_sort Zhang, Yuxun
collection PubMed
description 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 TFPα subunit. To test the effects of acylation on substate channeling through TFPα, 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 in vitro. 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.
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spelling pubmed-85138712021-10-14 The enzyme activity of mitochondrial trifunctional protein is not altered by lysine acetylation or lysine succinylation Zhang, Yuxun Goetzman, Eric PLoS One Research Article 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 TFPα subunit. To test the effects of acylation on substate channeling through TFPα, 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 in vitro. 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. Public Library of Science 2021-10-13 /pmc/articles/PMC8513871/ /pubmed/34644302 http://dx.doi.org/10.1371/journal.pone.0256619 Text en © 2021 Zhang, Goetzman https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Zhang, Yuxun
Goetzman, Eric
The enzyme activity of mitochondrial trifunctional protein is not altered by lysine acetylation or lysine succinylation
title The enzyme activity of mitochondrial trifunctional protein is not altered by lysine acetylation or lysine succinylation
title_full The enzyme activity of mitochondrial trifunctional protein is not altered by lysine acetylation or lysine succinylation
title_fullStr The enzyme activity of mitochondrial trifunctional protein is not altered by lysine acetylation or lysine succinylation
title_full_unstemmed The enzyme activity of mitochondrial trifunctional protein is not altered by lysine acetylation or lysine succinylation
title_short The enzyme activity of mitochondrial trifunctional protein is not altered by lysine acetylation or lysine succinylation
title_sort enzyme activity of mitochondrial trifunctional protein is not altered by lysine acetylation or lysine succinylation
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8513871/
https://www.ncbi.nlm.nih.gov/pubmed/34644302
http://dx.doi.org/10.1371/journal.pone.0256619
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