Analyzing protein dynamics using hydrogen exchange mass spectrometry

• Verfasst von: Hentze, Nikolai [VerfasserIn]
• Verfasst von: Mayer, Matthias P. [VerfasserIn]
• Titel: Analyzing protein dynamics using hydrogen exchange mass spectrometry
• Verf.angabe: Nikolai Hentze and Matthias P. Mayer
• E-Jahr: 2013
• Jahr: November 29, 2013
• Umfang: 1 Online-Ressource (1 Videodatei, 11:04 min)
• Umfang: ?
• Illustrationen: farbig
• Fussnoten: Enthält auch Textversion ; Gesehen am 13.04.2021 ; Wissenschaftlicher Film. Deutschland. 2013
• Titel Quelle: Enthalten in: JoVE. Video journal
• Ort Quelle: [S.l.] : [s.n.], 2006
• Jahr Quelle: 2013
• Band/Heft Quelle: (2013), 81, Artikel-ID e50839
• ISSN Quelle: 1940-087X
• DOI: doi:10.3791/50839
• Datenträger: Online-Ressource
• Dokumenttyp: Film
• Sprache: eng
• K10plus-PPN: 1753966868

Abstract

All cellular processes depend on the functionality of proteins. Although the functionality of a given protein is the direct consequence of its unique amino acid sequence, it is only realized by the folding of the polypeptide chain into a single defined three-dimensional arrangement or more commonly into an ensemble of interconverting conformations. Investigating the connection between protein conformation and its function is therefore essential for a complete understanding of how proteins are able to fulfill their great variety of tasks. One possibility to study conformational changes a protein undergoes while progressing through its functional cycle is hydrogen-1H/2H-exchange in combination with high-resolution mass spectrometry (HX-MS). HX-MS is a versatile and robust method that adds a new dimension to structural information obtained by e.g. crystallography. It is used to study protein folding and unfolding, binding of small molecule ligands, protein-protein interactions, conformational changes linked to enzyme catalysis, and allostery. In addition, HX-MS is often used when the amount of protein is very limited or crystallization of the protein is not feasible. Here we provide a general protocol for studying protein dynamics with HX-MS and describe as an example how to reveal the interaction interface of two proteins in a complex.