Evidence for an Allosteric S-Nitrosoglutathione Binding Site in S-Nitrosoglutathione Reductase (GSNOR)
| Autor: | Cristina Lento, James W. Gauld, Kathleen Fontana, Bulent Mutus, Leslie Ventimiglia, Derek J. Wilson, Sahar Nikoo, Nneamaka Onukwue, Bei-Lei Sun |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2019 |
| Předmět: |
0301 basic medicine
Physiology GSNO binding GSNO Clinical Biochemistry Allosteric regulation S-nitrosoglutathione reductase allosteric site for S-nitrosoglutathione Reductase 01 natural sciences Biochemistry Article 03 medical and health sciences S-nitrosoglutathione binding docking and molecular dynamics simulations hydrogen–deuterium exchange mass spectroscopy Molecular Biology chemistry.chemical_classification GSNOR 010405 organic chemistry Cell Biology 0104 chemical sciences 3. Good health 030104 developmental biology Enzyme chemistry Docking (molecular) Biophysics Hydrogen–deuterium exchange |
| Zdroj: | Antioxidants Volume 8 Issue 11 |
| ISSN: | 2076-3921 |
| DOI: | 10.3390/antiox8110545 |
| Popis: | Current research has identified S-nitrosoglutathione reductase (GSNOR) as the central enzyme for regulating protein S-nitrosylation. In addition, the dysregulation of GSNOR expression is implicated in several organ system pathologies including respiratory, cardiovascular, hematologic, and neurologic, making GSNOR a primary target for pharmacological intervention. This study demonstrates the kinetic activation of GSNOR by its substrate S-nitrosoglutathione (GSNO). GSNOR kinetic analysis data resulted in nonhyperbolic behavior that was successfully accommodated by the Hill&ndash Langmuir equation with a Hill coefficient of +1.75, indicating that the substrate, GSNO, was acting as a positive allosteric affector. Docking and molecular dynamics simulations were used to predict the location of the GSNO allosteric domain comprising the residues Asn185, Lys188, Gly321, and Lys323 in the vicinity of the structural Zn2+-binding site. GSNO binding to Lys188, Gly321, and Lys323 was further supported by hydrogen&ndash deuterium exchange mass spectroscopy (HDXMS), as deuterium exchange significantly decreased at these residues in the presence of GSNO. The site-directed mutagenesis of Lys188Ala and Lys323Ala resulted in the loss of allosteric behavior. Ultimately, this work unambiguously demonstrates that GSNO at large concentrations activates GSNOR by binding to an allosteric site comprised of the residues Asn185, Lys188, Gly321, and Lys323. The identification of an allosteric GSNO-binding domain on GSNOR is significant, as it provides a platform for pharmacological intervention to modulate the activity of this essential enzyme. |
| Databáze: | OpenAIRE |
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