Local molecular motions encode time-resolved infrared spectra of proteins
Time-resolved infrared spectroscopy probes protein dynamics over timescales spanning more than ten orders of magnitude, yet the molecular motions underlying the observed kinetic signatures have remained elusive. Here we combine transient infrared spectroscopy with nonequilibrium molecular dynamics simulations to establish a direct connection between experimental relaxation times and local structural motions. Studying single-domain allosteric proteins, we find that inter-residue contact distances provide the structural representation that most faithfully reproduces the experimental dynamics. Correlation analysis identifies localized networks of coordinated contacts that mediate communication between secondary-structure elements. The characteristic timescales of these contact networks quantitatively match the experimentally observed relaxation processes, enabling each kinetic step to be assigned to a specific molecular motion. Applied to allosteric signal propagation in PDZ3 and photoinduced ligand unbinding in PDZ2, this framework provides an atomistic picture of hierarchical protein relaxation and establishes a general framework for connecting transient infrared spectroscopy with the molecular mechanisms of protein dynamics.