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Hydrogen-Deuterium Exchange Coupled to Top- and Middle-Down Mass Spectrometry Reveals Histone Tail Dynamics before and after Nucleosome Assembly.
Karch, Kelly R; Coradin, Mariel; Zandarashvili, Levani; Kan, Zhong-Yuan; Gerace, Morgan; Englander, S Walter; Black, Ben E; Garcia, Benjamin A.
Afiliación
  • Karch KR; Department of Biochemistry and Molecular Biophysics, Perelman School of Medicine, University of Pennsylvania, Philadelphia 19104, USA; Epigenetics Institute, University of Pennsylvania, Philadelphia 19104, USA.
  • Coradin M; Department of Biochemistry and Molecular Biophysics, Perelman School of Medicine, University of Pennsylvania, Philadelphia 19104, USA; Epigenetics Institute, University of Pennsylvania, Philadelphia 19104, USA.
  • Zandarashvili L; Department of Biochemistry and Molecular Biophysics, Perelman School of Medicine, University of Pennsylvania, Philadelphia 19104, USA; Epigenetics Institute, University of Pennsylvania, Philadelphia 19104, USA.
  • Kan ZY; Department of Biochemistry and Molecular Biophysics, Perelman School of Medicine, University of Pennsylvania, Philadelphia 19104, USA.
  • Gerace M; Department of Biochemistry and Molecular Biophysics, Perelman School of Medicine, University of Pennsylvania, Philadelphia 19104, USA; Epigenetics Institute, University of Pennsylvania, Philadelphia 19104, USA.
  • Englander SW; Department of Biochemistry and Molecular Biophysics, Perelman School of Medicine, University of Pennsylvania, Philadelphia 19104, USA.
  • Black BE; Department of Biochemistry and Molecular Biophysics, Perelman School of Medicine, University of Pennsylvania, Philadelphia 19104, USA; Epigenetics Institute, University of Pennsylvania, Philadelphia 19104, USA.
  • Garcia BA; Department of Biochemistry and Molecular Biophysics, Perelman School of Medicine, University of Pennsylvania, Philadelphia 19104, USA; Epigenetics Institute, University of Pennsylvania, Philadelphia 19104, USA. Electronic address: bgarci@pennmedicine.upenn.edu.
Structure ; 26(12): 1651-1663.e3, 2018 12 04.
Article en En | MEDLINE | ID: mdl-30293810
Until recently, a major limitation of hydrogen-deuterium exchange mass spectrometry (HDX-MS) was that resolution of deuterium localization was limited to the length of the peptide generated during proteolysis. However, electron transfer dissociation (ETD) has been shown to preserve deuterium label in the gas phase, enabling better resolution. To date, this technology remains mostly limited to small, already well-characterized proteins. Here, we optimize, expand, and adapt HDX-MS tandem MS (MS/MS) capabilities to accommodate histone and nucleosomal complexes on top-down HDX-MS/MS and middle-down HDX-MS/MS platforms and demonstrate that near site-specific resolution of deuterium localization can be obtained with high reproducibility. We are able to study histone tail dynamics in unprecedented detail, which have evaded analysis by traditional structural biology techniques for decades, revealing important insights into chromatin biology. Together, the results of these studies highlight the versatility, reliability, and reproducibility of ETD-based HDX-MS/MS methodology to interrogate large protein and protein/DNA complexes.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Histonas / Nucleosomas Idioma: En Revista: Structure Asunto de la revista: BIOLOGIA MOLECULAR / BIOQUIMICA / BIOTECNOLOGIA Año: 2018 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Estados Unidos

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Histonas / Nucleosomas Idioma: En Revista: Structure Asunto de la revista: BIOLOGIA MOLECULAR / BIOQUIMICA / BIOTECNOLOGIA Año: 2018 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Estados Unidos