Fluorescence hole-burning and site-selective studies of LHCII

We report the observation of two types of changes in fluorescence spectra of LHCII at 4.2 K following intense illumination of the sample with a spectrally narrow laser beam at wavelengths between 678 and 686 nm. Nonspecific changes (burning-wavelength independent) are characterized by two relatively broad bands: a positive one at similar to 678.7 nm and a negative one at similar to 680.8 nm. These changes reveal a similar to 1.3-nm blue shift of the distribution of final emitters in LHCII, from 680.3 nm to similar to 679.0 nm independent of the excitation wavelength. Specific fluorescence changes (burning-wavelength dependent) are characterized by a sharp hole exactly at the burning wavelength, and positive changes directly to the shorter-and longer-wavelength side of the narrow hole. The negative changes are interpreted as zero-phonon holes, while the positive features are assigned to non-photochemical products. In the low-burning intensity experiment, in addition to the zero-phonon holes, we observed also the holes to the longer wavelength of the zero-phonon hole, which were assigned to a sum of phonon and pseudo-phonon side bands. The shapes of these extra holes are identical to the shapes of the holes revealed in the fluorescence line narrowing experiment. On the basis of the low-burning intensity experiment we estimated the upper limit of the electron-phonon coupling strength for LHCII, characterized by a Huang-Rhys factor of 1.5.