Regularities of Bonding of Chlorin e 6 to the Oligomeric Enzyme Lactate Dehydrogenase

Using differential-spectrophotometry, spectral-luminescence, and polarization methods, we have investigated regularities of complexing of a promising photodynamic sensitizer — chlorin e₆ — with a key glycolytic enzyme — lactate dehydrogenase (LDH). The parameters of the dye–enzyme complex have been estimated by the difference between the spectral characteristics of the free dye and the dye bonded to the enzyme. It is shown that the tetrameric LDH molecule forms an equilibrium complex with four chlorin molecules and the sensitizer is bonded independently to each subunit entering into the composition of the tetramer. It has been established that the spectral characteristics of chlorin bonded to LDH are sensitive to the structure transformations arising in the active center of the enzyme as a result of the formation of an unproductive enzyme–coenzyme–substrate complex, which allows the conclusion that the dye is localized in the neighborhood of the active center of LDH.