title: Analyzing protein dynamics using hydrogen exchange mass spectrometry.
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-(1)H/(2)H-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.
authors: Zentrum für Molekulare Biologie der Universität Heidelberg (ZMBH), University of Heidelberg.
authors: Nikolai
authors: N
authors: Hentze
authors: Matthias P
authors: MP
authors: Mayer
mesh_terms: D041961:Deuterium Exchange Measurement / Q000379:methods*; D007700:Kinetics; D013058:Mass Spectrometry / Q000379:methods*; D011487:Protein Conformation; D017510:Protein Folding; D054730:Protein Interaction Domains and Motifs; D011506:Proteins / Q000737:chemistry*
chemical_list: D011506:Proteins