Inhibitory Investigations of Acyl-CoA Derivatives against Human Lipoxygenase Isozymes

• Published in: International journal of molecular sciences Vol. 24; no. 13; p. 10941
• Main Authors: Tran, Michelle, Yang, Kevin, Glukhova, Alisa, Holinstat, Michael, Holman, Theodore
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 30.06.2023; MDPI
• Subjects: Acyl Coenzyme A - metabolism; acyl-coenzyme A; Allosteric properties; Arachidonate 12-lipoxygenase; Coenzyme A; Enzymes; Ethylenediaminetetraacetic acid; Fatty acids; Humans; Inflammation; inhibition; Investigations; Isoenzymes; Linoleic acid; Lipid metabolism; Lipids; Lipoxygenase; Liquid oxygen; LOX-1 protein; Metabolism; Oxylipins; Physiological aspects; Resveratrol; Reticulocytes; Scavenger Receptors, Class E; Substrate inhibition; Type 2 diabetes; Australia; acyl-coenzyme A; lipoxygenase; inhibition
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms241310941

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 = 32 μM) and h15-LOX-2 (IC = 0.62 μM), stearoyl-CoA against h15-LOX-1 (IC = 4.2 μM), and palmitoleoyl-CoA against h5-LOX (IC = 2.0 μM). The inhibition of h15-LOX-2 by oleoyl-CoA was further determined to be allosteric inhibition with a 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 ( = 7.5 +/- 0.4 s , = 0.62 +/- 0.1 µM s ). 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.