Neuroglobin Provides a Convenient Scaffold to Investigate the Triplet-State Properties of Porphyrins by Time-Resolved EPR Spectroscopy and Magnetophotoselection

The photo-excited triplet state of Zn-protoporphyrin IX located in the heme pocket of human neuroglobin has been investigated by time-resolved EPR coupled to magnetophotoselection. The triplet state in the protein matrix has been compared with the model complex in organic glass, considering both non-coordinating and coordinating solvent mixtures. The protein matrix plays an important role in stabilizing the coordination of the embedded chromophore, resulting in a more homogeneous environment relative to that of the chromophore in a glassy solvent, even in the presence of an axial nitrogenous ligand like pyridine. The EPR spectral parameters point out a slow Jahn–Teller interconversion between slightly different triplet states, both in organic solvent and in the protein matrix. The EPR-magnetophotoselection allows us to propose a reinterpretation of the assignment of the Q bands in the electronic absorption spectrum.

Products of Docosahexaenoate Oxidation as Contributors to Photosensitising Properties of Retinal Lipofuscin

Retinal lipofuscin which accumulates with age in the retinal pigment epithelium (RPE) is subjected to daily exposures to high fluxes of visible light and exhibits potent photosensitising properties; however, the molecules responsible for its photoreactivity remain unknown. Here, we demonstrate that autooxidation of docosahexaenoate (DHE) leads to the formation of products absorbing, in addition to UVB and UVA light, also visible light. The products of DHE oxidation exhibit potent photosensitising properties similar to photosensitising properties of lipofuscin, including generation of an excited triplet state with similar characteristics as the lipofuscin triplet state, and photosensitised formation of singlet oxygen and superoxide. The quantum yields of singlet oxygen and superoxide generation by oxidised DHE photoexcited with visible light are 2.4- and 3.6-fold higher, respectively, than for lipofuscin, which is consistent with the fact that lipofuscin contains some chromophores which do contribute to the absorption of light but not so much to its photosensitising properties. Importantly, the wavelength dependence of photooxidation induced by DHE oxidation products normalised to equal numbers of incident photons is also similar to that of lipofuscin—it steeply increases with decreasing wavelength. Altogether, our results demonstrate that products of DHE oxidation include potent photosensitiser(s) which are likely to contribute to lipofuscin photoreactivity.

Saturation in the principal (excited) channel of four-level medium as an effective two-level configuration

Saturation of complex molecules affects the nonlinear processes occurring in the medium. The medium can be described by two-, three-, and four-level configurations. The principal singlet-singlet and excited triplet-triplet channels of a four-level, N-type, configuration were used to study a nonlinear medium, which was excited by radiation with two frequencies. Under weak population of energy levels in the triplet-triplet channel (a small ratio of the total probability of spontaneous and irradiate transitions for both channels p23/p31), the principal (singlet-singlet) channel is similar to a two-level configuration. At the same time, the effective population of the energy levels in the triplet-triplet channel (the ratio p23/p31 is large and the radiation intensity in the principal channel is high) makes the excited (triplet-triplet) channel coincide with the two-level configuration. The saturation intensity of radiation in principal (excited) channel, as effective two-level configuration, and nonlinear processes in this channel can be controlled by external radiation acting on excited (principal) channel. Finally, the average excitation of molecules into the energy levels in the triplet-triplet channel (ratio p23/p31 ≈ 1) makes the four-level configuration similar to the three-level configuration with effective level as a result of combination between second singlet level S2 and first triplet level T1.