Small molecule-induced oxidation of protein disulfide isomerase is neuroprotective
| العنوان: | Small molecule-induced oxidation of protein disulfide isomerase is neuroprotective |
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| المؤلفون: | Michael M. Gaschler, Lewis M. Brown, Brent R. Stockwell, Ryan M. Colligan, Donald C. Lo, Denise E. Dunn, Anna Kaplan, Arthur G. Palmer |
| المصدر: | Proceedings of the National Academy of Sciences. 112 |
| بيانات النشر: | Proceedings of the National Academy of Sciences, 2015. |
| سنة النشر: | 2015 |
| مصطلحات موضوعية: | inorganic chemicals, Protein Disulfide-Isomerases, Nerve Tissue Proteins, PC12 Cells, Mice, Catalytic Domain, Animals, Enzyme Inhibitors, Binding site, Protein disulfide-isomerase, Nuclear Magnetic Resonance, Biomolecular, Huntingtin Protein, Multidisciplinary, Chemistry, Endoplasmic reticulum, Isothermal titration calorimetry, Small molecule, Corpus Striatum, Rats, nervous system diseases, body regions, Neuroprotective Agents, PNAS Plus, Biochemistry, Microsomes, Liver, Microsome, Protein folding, Oxidation-Reduction, Heteronuclear single quantum coherence spectroscopy |
| الوصف: | Protein disulfide isomerase (PDI) is a chaperone protein in the endoplasmic reticulum that is up-regulated in mouse models of, and brains of patients with, neurodegenerative diseases involving protein misfolding. PDI's role in these diseases, however, is not fully understood. Here, we report the discovery of a reversible, neuroprotective lead optimized compound (LOC)14, that acts as a modulator of PDI. LOC14 was identified using a high-throughput screen of ∼10,000 lead-optimized compounds for potent rescue of viability of PC12 cells expressing mutant huntingtin protein, followed by an evaluation of compounds on PDI reductase activity in an in vitro screen. Isothermal titration calorimetry and fluorescence experiments revealed that binding to PDI was reversible with a Kd of 62 nM, suggesting LOC14 to be the most potent PDI inhibitor reported to date. Using 2D heteronuclear single quantum correlation NMR experiments, we were able to map the binding site of LOC14 as being adjacent to the active site and to observe that binding of LOC14 forces PDI to adopt an oxidized conformation. Furthermore, we found that LOC14-induced oxidation of PDI has a neuroprotective effect not only in cell culture, but also in corticostriatal brain slice cultures. LOC14 exhibited high stability in mouse liver microsomes and blood plasma, low intrinsic microsome clearance, and low plasma-protein binding. These results suggest that LOC14 is a promising lead compound to evaluate the potential therapeutic effects of modulating PDI in animal models of disease. |
| تدمد: | 1091-6490 0027-8424 |
| DOI: | 10.1073/pnas.1500439112 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::b8f3a55916504b11b44f80754b0d8517 https://doi.org/10.1073/pnas.1500439112 |
| Rights: | OPEN |
| رقم الانضمام: | edsair.doi.dedup.....b8f3a55916504b11b44f80754b0d8517 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 10916490 00278424 |
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| DOI: | 10.1073/pnas.1500439112 |