Revealing the dynamic allosteric changes required for formation of the cysteine synthase complex by hydrogen-deuterium exchange MS
| Autor: | Stefano Bettati, Eleanor R. Dickinson, Barbara Pioselli, Marialaura Marchetti, Barbara Campanini, Brenda Rosa, Kasper D. Rand |
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| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
BE
back exchange Stereochemistry Allosteric regulation Protein Data Bank (RCSB PDB) Trimer protein–protein interactions Cysteine synthase Biochemistry Mass Spectrometry Analytical Chemistry Protein–protein interaction chemistry.chemical_compound PDB Protein Data Bank Molecular Biology cysteine biosynthesis Cysteine Synthase CS cysteine synthase biology Research HDX-MS hydrogen/deuterium exchange MS SAXS small-angle X-ray scattering Deuterium Exchange Measurement Deuterium bienzyme complex Reaxys Chemistry database information allosteric regulation CI confidence interval chemistry OAS O-acetylserine biology.protein DDA data-dependent acquisition Cysteine synthase complex O-Acetylserine Cysteine Chromatography Liquid Hydrogen |
| Zdroj: | Molecular & cellular proteomics 20 (2021). doi:10.1016/j.mcpro.2021.100098 info:cnr-pdr/source/autori:Rosa B.; Dickinson E.R.; Marchetti M.; Campanini B.; Pioselli B.; Bettati S.; Rand K.D./titolo:Revealing the dynamic allosteric changes required for formation of the cysteine synthase complex by hydrogen-deuterium exchange MS/doi:10.1016%2Fj.mcpro.2021.100098/rivista:Molecular & cellular proteomics/anno:2021/pagina_da:/pagina_a:/intervallo_pagine:/volume:20 Molecular & Cellular Proteomics : MCP |
| DOI: | 10.1016/j.mcpro.2021.100098 |
| Popis: | CysE and CysK, the last two enzymes of the cysteine biosynthetic pathway, engage in a bienzyme complex, cysteine synthase, with yet incompletely characterized three-dimensional structure and regulatory function. Being absent in mammals, the two enzymes and their complex are attractive targets for antibacterial drugs. We have used hydrogen/deuterium exchange MS to unveil how complex formation affects the conformational dynamics of CysK and CysE. Our results support a model where CysE is present in solution as a dimer of trimers, and each trimer can bind one CysK homodimer. When CysK binds to one CysE monomer, intratrimer allosteric communication ensures conformational and dynamic symmetry within the trimer. Furthermore, a long-range allosteric signal propagates through CysE to induce stabilization of the interface between the two CysE trimers, preparing the second trimer for binding the second CysK with a nonrandom orientation. These results provide new molecular insights into the allosteric formation of the cysteine synthase complex and could help guide antibacterial drug design. Graphical Abstract Highlights • HDX-MS reveals complex formation impact on conformational dynamics of CysK and CysE. • CysK binding ensures conformational symmetry within the CysE trimer. • Long-range allostery propagates through CysE stabilizing the inter-trimer interface. • Insights into the allostery of CS complex could help guide antibacterial drug design. In Brief We have used hydrogen/deuterium exchange MS to unveil the allosteric changes occurring during complex formation of CysE and CysK, the last two enzymes of cysteine biosynthetic pathway in bacteria. Significant changes in conformation and dynamics occur in each protein upon complex formation, including long distance intraprotein and interprotein communication. Being absent in mammals, both CysE and CysK represent potential targets for new antibacterial drugs, and our results on the formation and allostery of the complex could help guide drug development. |
| Databáze: | OpenAIRE |
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