SEC-SAXS and HDX-MS: A powerful combination. The case of the calcium-binding domain of a bacterial toxin
| Autor: | O'Brien, Darragh, Brier, Sebastien, Ladant, Daniel, Durand, Dominique, Chenal, Alexandre, Vachette, Patrice |
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| Přispěvatelé: | Biochimie des Interactions Macromoléculaires / Biochemistry of Macromolecular Interactions, Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Centre de Recherche et Innovation Technologique (CITECH), Institut Pasteur [Paris] (IP), Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Fonction et Architecture des Assemblages Macromoléculaires (FAAM), Département Biochimie, Biophysique et Biologie Structurale (B3S), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), This work was supported by CNRS, Institut Pasteur and the program PasteurInnoV., Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Institut Pasteur [Paris] |
| Jazyk: | angličtina |
| Rok vydání: | 2018 |
| Předmět: |
Models
Molecular MESH: Adenylate Cyclase Toxin/chemistry [SDV]Life Sciences [q-bio] RTX motif MESH: Calcium/chemistry hydrogen/deuterium exchange mass spectrometry Bordetella pertussis Mass Spectrometry X-Ray Diffraction Scattering Small Angle [SDV.BBM]Life Sciences [q-bio]/Biochemistry Molecular Biology MESH: Scattering Small Angle MESH: Mass Spectrometry Binding Sites calcium MESH: X-Ray Diffraction Deuterium Exchange Measurement size-exclusion chromatography MESH: Binding Sites MESH: Deuterium Exchange Measurement small-angle X-ray scattering Adenylate Cyclase Toxin Quantum Theory MESH: Quantum Theory MESH: Models Molecular MESH: Bordetella pertussis/chemistry |
| Zdroj: | Biotechnology and Applied Biochemistry Biotechnology and Applied Biochemistry, 2018, 65 (1), pp.62-68. ⟨10.1002/bab.1577⟩ Biotechnology and Applied Biochemistry, Wiley, 2018, 65 (1), pp.62-68. ⟨10.1002/bab.1577⟩ |
| ISSN: | 0885-4513 1470-8744 |
| Popis: | International audience; Small-angle X-ray scattering (SAXS) is a relatively simple experimental technique that provides information on the global conformation of macromolecules in solution, be they fully structured, partially, or extensively unfolded. Size exclusion chromatography in line with a SAXS measuring cell considerably improves the monodispersity and ideality of solutions, the two main requirements of a "good" SAXS sample. Hydrogen/deuterium exchange monitored by mass spectrometry (HDX-MS) offers a wealth of information regarding the solvent accessibility at the local (peptide) level. It constitutes a sensitive probe of local flexibility and, more generally, of structural dynamics. The combination of both approaches presented here is very powerful, as illustrated by the case of RD, a calcium-binding protein that is part of a bacterial virulence factor. |
| Databáze: | OpenAIRE |
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