CIDEP Created by the Quenching of Photo-Excited Tryptophan at Protein Surface: A Challenge to CIDEP Probing of Protein Structural Changes

Quenching of the triplet excited state of molecular tryptophan by nitroxide radical in 1,4-dioxane and water solutions was investigated by means of time-resolved electron paramagnetic resonance (EPR) and Fourier-transform (FT)-EPR. The chemically induced dynamic electron polarization (CIDEP) signals with net emissive phase were recorded at these quenching events and were analyzed through radical-triplet pair mechanism. The CIDEP time profiles were well reproduced by Bloch and kinetic equations, assuming radical-triplet pair mechanism with the appropriate quenching rate constants. From a comparison of the simulation and the experiment, CIDEP enhancement factor in 1,4-dioxane was determined to be −30 × P _eq, where P _eq is the spin polarization of nitroxide at thermal equilibrium. Net emissive CIDEP was also observed by FT-EPR measurements on the nitroxide quenching of the triplet excited state of tryptophan residue in α-lactalbumin. Magnitude of CIDEP created in α-lactalbumin/nitroxide system depends on the pH condition of α-lactalbumin solution, which is related to protein folding dynamics. We argue the CIDEP mechanism at the α-lactalbumin surface and propose a possibility of a novel CIDEP method to probe a protein surface and structural changes.