Abstract
Using time-resolved step-scan FTIR spectroscopy, it has become possible to identify two
different M states in the 5 to 300 μs time range, which clearly differ from the MN state.
The identification has become possible by measuring a pure BR→KL difference
spectrum at 100 ns, which can be used to correct for the contributions of this
intermediate in the later difference spectra. The subtraction of the later corrected
difference spectra thus represent L→M1, M2 difference spectra with varying relative
amount of the two M states. The comparison of these spectra clearly reveals differences
in the amide-II spectral range, and possibly also in the amide-I range. From these
observations it can be concluded that the two M states do not differ in the chromophore
structure but in the protein structure.