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Inhibitory Investigations of Acyl-CoA Derivatives against Human Lipoxygenase Isozymes
Lipid metabolism is a complex process crucial for energy production resulting in high levels of acyl-coenzyme A (acyl-CoA) molecules in the cell. Acyl-CoAs have also been implicated in inflammation, which could be possibly linked to lipoxygenase (LOX) biochemistry by the observation that an acyl-CoA...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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MDPI
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10341549/ https://www.ncbi.nlm.nih.gov/pubmed/37446119 http://dx.doi.org/10.3390/ijms241310941 |
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author | Tran, Michelle Yang, Kevin Glukhova, Alisa Holinstat, Michael Holman, Theodore |
author_facet | Tran, Michelle Yang, Kevin Glukhova, Alisa Holinstat, Michael Holman, Theodore |
author_sort | Tran, Michelle |
collection | PubMed |
description | Lipid metabolism is a complex process crucial for energy production resulting in high levels of acyl-coenzyme A (acyl-CoA) molecules in the cell. Acyl-CoAs have also been implicated in inflammation, which could be possibly linked to lipoxygenase (LOX) biochemistry by the observation that an acyl-CoA was bound to human platelet 12-lipoxygenase via cryo-EM. Given that LOX isozymes play a pivotal role in inflammation, a more thorough investigation of the inhibitory effects of acyl-CoAs on lipoxygenase isozymes was judged to be warranted. Subsequently, it was determined that C18 acyl-CoA derivatives were the most potent against h12-LOX, human reticulocyte 15-LOX-1 (h15-LOX-1), and human endothelial 15-LOX-2 (h15-LOX-2), while C16 acyl-CoAs were more potent against human 5-LOX. Specifically, oleoyl-CoA (18:1) was most potent against h12-LOX (IC(50) = 32 μM) and h15-LOX-2 (IC(50) = 0.62 μM), stearoyl-CoA against h15-LOX-1 (IC(50) = 4.2 μM), and palmitoleoyl-CoA against h5-LOX (IC(50) = 2.0 μM). The inhibition of h15-LOX-2 by oleoyl-CoA was further determined to be allosteric inhibition with a K(i) of 82 +/− 70 nM, an α of 3.2 +/− 1, a β of 0.30 +/− 0.07, and a β/α = 0.09. Interestingly, linoleoyl-CoA (18:2) was a weak inhibitor against h5-LOX, h12-LOX, and h15-LOX-1 but a rapid substrate for h15-LOX-1, with comparable kinetic rates to free linoleic acid (k(cat) = 7.5 +/− 0.4 s(−1), k(cat)/K(M) = 0.62 +/− 0.1 µM(−1)s(−1)). Additionally, it was determined that methylated fatty acids were not substrates but rather weak inhibitors. These findings imply a greater role for acyl-CoAs in the regulation of LOX activity in the cell, either through inhibition of novel oxylipin species or as a novel source of oxylipin-CoAs. |
format | Online Article Text |
id | pubmed-10341549 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-103415492023-07-14 Inhibitory Investigations of Acyl-CoA Derivatives against Human Lipoxygenase Isozymes Tran, Michelle Yang, Kevin Glukhova, Alisa Holinstat, Michael Holman, Theodore Int J Mol Sci Article Lipid metabolism is a complex process crucial for energy production resulting in high levels of acyl-coenzyme A (acyl-CoA) molecules in the cell. Acyl-CoAs have also been implicated in inflammation, which could be possibly linked to lipoxygenase (LOX) biochemistry by the observation that an acyl-CoA was bound to human platelet 12-lipoxygenase via cryo-EM. Given that LOX isozymes play a pivotal role in inflammation, a more thorough investigation of the inhibitory effects of acyl-CoAs on lipoxygenase isozymes was judged to be warranted. Subsequently, it was determined that C18 acyl-CoA derivatives were the most potent against h12-LOX, human reticulocyte 15-LOX-1 (h15-LOX-1), and human endothelial 15-LOX-2 (h15-LOX-2), while C16 acyl-CoAs were more potent against human 5-LOX. Specifically, oleoyl-CoA (18:1) was most potent against h12-LOX (IC(50) = 32 μM) and h15-LOX-2 (IC(50) = 0.62 μM), stearoyl-CoA against h15-LOX-1 (IC(50) = 4.2 μM), and palmitoleoyl-CoA against h5-LOX (IC(50) = 2.0 μM). The inhibition of h15-LOX-2 by oleoyl-CoA was further determined to be allosteric inhibition with a K(i) of 82 +/− 70 nM, an α of 3.2 +/− 1, a β of 0.30 +/− 0.07, and a β/α = 0.09. Interestingly, linoleoyl-CoA (18:2) was a weak inhibitor against h5-LOX, h12-LOX, and h15-LOX-1 but a rapid substrate for h15-LOX-1, with comparable kinetic rates to free linoleic acid (k(cat) = 7.5 +/− 0.4 s(−1), k(cat)/K(M) = 0.62 +/− 0.1 µM(−1)s(−1)). Additionally, it was determined that methylated fatty acids were not substrates but rather weak inhibitors. These findings imply a greater role for acyl-CoAs in the regulation of LOX activity in the cell, either through inhibition of novel oxylipin species or as a novel source of oxylipin-CoAs. MDPI 2023-06-30 /pmc/articles/PMC10341549/ /pubmed/37446119 http://dx.doi.org/10.3390/ijms241310941 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Tran, Michelle Yang, Kevin Glukhova, Alisa Holinstat, Michael Holman, Theodore Inhibitory Investigations of Acyl-CoA Derivatives against Human Lipoxygenase Isozymes |
title | Inhibitory Investigations of Acyl-CoA Derivatives against Human Lipoxygenase Isozymes |
title_full | Inhibitory Investigations of Acyl-CoA Derivatives against Human Lipoxygenase Isozymes |
title_fullStr | Inhibitory Investigations of Acyl-CoA Derivatives against Human Lipoxygenase Isozymes |
title_full_unstemmed | Inhibitory Investigations of Acyl-CoA Derivatives against Human Lipoxygenase Isozymes |
title_short | Inhibitory Investigations of Acyl-CoA Derivatives against Human Lipoxygenase Isozymes |
title_sort | inhibitory investigations of acyl-coa derivatives against human lipoxygenase isozymes |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10341549/ https://www.ncbi.nlm.nih.gov/pubmed/37446119 http://dx.doi.org/10.3390/ijms241310941 |
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