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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...
Autores principales: | , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2021
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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. |
format | Online Article Text |
id | pubmed-8513871 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
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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