Protocol for determining protein dynamics using FT-IR spectroscopy
Hao Shen, Cuiping Fu, Junting Zhang, Bin Feng, Shaoning Yu
Abstract
In biological systems, protein function depends on spatial and temporal changes known as protein dynamics, which can be probed by amide hydrogen/deuterium (H/D) exchange. Here, we present a protocol for determining protein dynamics by Fourier-transform infrared (FT-IR) spectroscopy. We describe steps for protein sample preparation and FT-IR spectra collection. We then detail procedures for spectra analysis. Applications include the effects of protein mutation or protein and metal ion or ligand interactions on the protein H/D exchange rate. For complete details on the use and execution of this protocol, please refer to Yu et al. (2013).1
Topics & Concepts
Protein dynamicsInfrared spectroscopySpectroscopyChemistryHydrogen–deuterium exchangeProtein functionDynamics (music)DeuteriumProtein structureMolecular dynamicsAnalytical Chemistry (journal)Chemical physicsBiological systemComputational chemistryPhysicsBiologyBiochemistryAtomic physicsChromatographyOrganic chemistryGeneAcousticsQuantum mechanicsMass Spectrometry Techniques and ApplicationsMetabolomics and Mass Spectrometry StudiesProtein Structure and Dynamics