The Mitochondrial Protein MitoNEET as a Probe for the Allostery of Glutamate Dehydrogenase
The proteins glutamate dehydrogenase (GDH) and mitoNEET are both targets of drug development efforts to treat metabolic disorders, cancer, and neurodegenerative diseases. However, these two proteins differ starkly in the current knowledge about ligand binding sites. MitoNEET is a [2Fe-2S]-containing...
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MDPI AG
2022-11-01
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| Series: | Molecules |
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| Online Access: | https://www.mdpi.com/1420-3049/27/23/8314 |
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| author | Chimere Nnatubeugo Erica Johnson Sarah Gisondi Felicia Roland Werner J. Geldenhuys Michael A. Menze Mary E. Konkle |
| author_facet | Chimere Nnatubeugo Erica Johnson Sarah Gisondi Felicia Roland Werner J. Geldenhuys Michael A. Menze Mary E. Konkle |
| author_sort | Chimere Nnatubeugo |
| collection | DOAJ |
| description | The proteins glutamate dehydrogenase (GDH) and mitoNEET are both targets of drug development efforts to treat metabolic disorders, cancer, and neurodegenerative diseases. However, these two proteins differ starkly in the current knowledge about ligand binding sites. MitoNEET is a [2Fe-2S]-containing protein with no obvious binding site for small ligands observed in its crystal structures. In contrast, GDH is known to have a variety of ligands at multiple allosteric sites thereby leading to complex regulation in activity. In fact, while GDH can utilize either NAD(H) or NADP(H) for catalysis at the active site, only NAD(H) binds at a regulatory site to inhibit GDH activity. Previously, we found that mitoNEET forms a covalent bond with GDH in vitro and increases the catalytic activity of the enzyme. In this study we evaluated the effects of mitoNEET binding on the allosteric control of GDH conferred by inhibitors. We examined all effectors using NAD or NADP as the coenzyme to determine allosteric linkage by the NAD-binding regulatory site. We found that GDH activity, in the presence of the inhibitory palmitoyl-CoA and EGCG, can be rescued by mitoNEET, regardless of the coenzyme used. This suggests that mitoNEET rescues GDH by stabilizing the open conformation. |
| first_indexed | 2024-03-09T17:39:22Z |
| format | Article |
| id | doaj.art-169724906be7449092119ccba5ac0da7 |
| institution | Directory Open Access Journal |
| issn | 1420-3049 |
| language | English |
| last_indexed | 2024-03-09T17:39:22Z |
| publishDate | 2022-11-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Molecules |
| spelling | doaj.art-169724906be7449092119ccba5ac0da72023-11-24T11:40:12ZengMDPI AGMolecules1420-30492022-11-012723831410.3390/molecules27238314The Mitochondrial Protein MitoNEET as a Probe for the Allostery of Glutamate DehydrogenaseChimere Nnatubeugo0Erica Johnson1Sarah Gisondi2Felicia Roland3Werner J. Geldenhuys4Michael A. Menze5Mary E. Konkle6Department of Chemistry, Ball State University, 2000 W. University Avenue, Muncie, IN 47306, USADepartment of Chemistry, Ball State University, 2000 W. University Avenue, Muncie, IN 47306, USADepartment of Chemistry, Eastern Illinois University, Charleston, IL 61920, USADepartment of Chemistry, Eastern Illinois University, Charleston, IL 61920, USAPharmaceutical Sciences, School of Pharmacy, West Virginia University, Morgantown, WV 26506, USADepartment of Biology, University of Louisville, Louisville, KY 40292, USADepartment of Chemistry, Ball State University, 2000 W. University Avenue, Muncie, IN 47306, USAThe proteins glutamate dehydrogenase (GDH) and mitoNEET are both targets of drug development efforts to treat metabolic disorders, cancer, and neurodegenerative diseases. However, these two proteins differ starkly in the current knowledge about ligand binding sites. MitoNEET is a [2Fe-2S]-containing protein with no obvious binding site for small ligands observed in its crystal structures. In contrast, GDH is known to have a variety of ligands at multiple allosteric sites thereby leading to complex regulation in activity. In fact, while GDH can utilize either NAD(H) or NADP(H) for catalysis at the active site, only NAD(H) binds at a regulatory site to inhibit GDH activity. Previously, we found that mitoNEET forms a covalent bond with GDH in vitro and increases the catalytic activity of the enzyme. In this study we evaluated the effects of mitoNEET binding on the allosteric control of GDH conferred by inhibitors. We examined all effectors using NAD or NADP as the coenzyme to determine allosteric linkage by the NAD-binding regulatory site. We found that GDH activity, in the presence of the inhibitory palmitoyl-CoA and EGCG, can be rescued by mitoNEET, regardless of the coenzyme used. This suggests that mitoNEET rescues GDH by stabilizing the open conformation.https://www.mdpi.com/1420-3049/27/23/8314free fatty acidsbeta-oxidationmitochondriagreen teahyperinsulinism |
| spellingShingle | Chimere Nnatubeugo Erica Johnson Sarah Gisondi Felicia Roland Werner J. Geldenhuys Michael A. Menze Mary E. Konkle The Mitochondrial Protein MitoNEET as a Probe for the Allostery of Glutamate Dehydrogenase Molecules free fatty acids beta-oxidation mitochondria green tea hyperinsulinism |
| title | The Mitochondrial Protein MitoNEET as a Probe for the Allostery of Glutamate Dehydrogenase |
| title_full | The Mitochondrial Protein MitoNEET as a Probe for the Allostery of Glutamate Dehydrogenase |
| title_fullStr | The Mitochondrial Protein MitoNEET as a Probe for the Allostery of Glutamate Dehydrogenase |
| title_full_unstemmed | The Mitochondrial Protein MitoNEET as a Probe for the Allostery of Glutamate Dehydrogenase |
| title_short | The Mitochondrial Protein MitoNEET as a Probe for the Allostery of Glutamate Dehydrogenase |
| title_sort | mitochondrial protein mitoneet as a probe for the allostery of glutamate dehydrogenase |
| topic | free fatty acids beta-oxidation mitochondria green tea hyperinsulinism |
| url | https://www.mdpi.com/1420-3049/27/23/8314 |
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