Abstract
The tyrosine ring mode is an intrinsic non-perturbing site-specific infrared reporter for conformational dynamics within protein systems. This transition is influenced by direct and indirect interactions associated with the electron-donating ability and the hydrophobicity of the surrounding molecules. Utilizing an intrinsic tyrosine moiety, two-dimensional infrared spectra of Trp-cage, often called the "hydrogen atom" of protein folding, were measured in the folded and denatured states to uncover the dynamics of the hydrophobic core. The vibrational lifetimes and the correlation decays of the tyrosine ring mode showed significant changes upon both temperature and chemical denaturation of the Trp-cage miniprotein, indicating important structural features of the hydrophobic core and its dynamics. The observed Trp6-Tyr3 interactions are in good agreement with the prior studies of the folded state, but they reach beyond the static structure. These stacking interactions and orientations fluctuate on the picosecond time scale as measured through the spectral dephasing within a dehydrated environment.