The allosteric interplay between S-nitrosylation and glycine binding controls the activity of human serine racemase
| Autor: | Marchesani, Gianquinto, Autiero, Michielon, Campanini, Faggiano, Bettati, Mozzarelli, Spyrakis, Bruno |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
0301 basic medicine
Protein Conformation Stereochemistry Allosteric regulation Racemases and Epimerases D-serine Biochemistry NMDA receptors 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine Glycine binding Allosteric Regulation Biosynthesis Catalytic Domain serine racemase Humans Molecular Biology pyridoxal phosphate Binding Sites allosteric modulation Chemistry Nitrosylation nitrosylation Cell Biology S-Nitrosylation fluorescence spectroscopy Ligand (biochemistry) glycine molecular dynamics Kinetics 030104 developmental biology 030220 oncology & carcinogenesis Serine racemase Glycine Oxidation-Reduction |
| Zdroj: | European journal of biochemistry 288 (2021): 3034–3054. doi:10.1111/febs.15645 info:cnr-pdr/source/autori:Marchesani, F. and Gianquinto, E. and Autiero, I. and Michielon, A. and Campanini, B. and Faggiano, S. and Bettati, S. and Mozzarelli, A. and Spyrakis, F. and Bruno, S./titolo:The allosteric interplay between S-nitrosylation and glycine binding controls the activity of human serine racemase/doi:10.1111%2Ffebs.15645/rivista:European journal of biochemistry/anno:2021/pagina_da:3034/pagina_a:3054/intervallo_pagine:3034–3054/volume:288 |
| DOI: | 10.1111/febs.15645 |
| Popis: | Human serine racemase (hSR) catalyzes the biosynthesis of D-serine, an obligatory co-agonist of the NMDA receptors. It was previously found that the reversible S-nitrosylation of Cys113 reduces hSR activity. Here, we show by site-directed mutagenesis, fluorescence spectroscopy, mass spectrometry, and molecular dynamics that S-nitrosylation stabilizes an open, less-active conformation of the enzyme. The reaction of hSR with either NO or nitroso donors is conformation-dependent and occurs only in the conformation stabilized by the allosteric effector ATP, in which the e-amino group of Lys114 acts as a base towards the thiol group of Cys113. In the closed conformation stabilized by glycine - an active-site ligand of hSR - the side chain of Lys114 moves away from that of Cys113, while the carboxyl side-chain group of Asp318 moves significantly closer, increasing the thiol pKa and preventing the reaction. We conclude that ATP binding, glycine binding, and S-nitrosylation constitute a three-way regulation mechanism for the tight control of hSR activity. We also show that Cys113 undergoes H2 O2 -mediated oxidation, with loss of enzyme activity, a reaction also dependent on hSR conformation. |
| Databáze: | OpenAIRE |
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