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Monday, January 13, 2014

Photodamage in a Mitochondrial Membrane Model Modulated by the Topology of Cationic and Anionic Meso-Tetrakis Porphyrin Free Bases

Cintia Kawai, Juliana C. Araújo-Chaves, Taciana Magrini, Camila O. C. C. Sanches, Sandra M. S. Pinto, Herculano Martinho, Nasser Daghastanli, Iseli L. Nantes

The photodynamic effects of the cationic TMPyP (meso-tetrakis [N-methyl-4-pyridyl]porphyrin) and the anionic TPPS4 (meso-tetrakis[4-sulfonatophenyl]porphyrin) against PC/CL phosphatidylcholine/cardiolipin (85/15%) membranes were probed to address the influence of phorphyrin binding on lipid damage. Electronic absorption spectroscopy and zeta potential demonstrated that only TMPyP binds to PC/CL large unilamellar vesicles (LUVs). The photodamage after irradiation with visible light was analyzed by dosages of lipid peroxides (LOOH) and thiobarbituric reactive substance and by a contrast phase image of the giant unillamelar vesicles (GUVs). Both TMPyP and TPPS4 promoted differentiated quantitative and qualitative damages on LUVs and GUVs. The damages were more extensive and faster by using the cationic porphyrin. The increase of LOOH was higher in the presence of D2O, and was impaired by sodium azide and sorbic acid. The effect of D2O was higher for TPPS4 as the photosensitizer. The use of DCFH demonstrated that liposomes prevent the photo-bleaching of TMPyP. The results are consistent with a more stable TMPyP that generates long-lived singlet oxygen preferentially partitioned in the bilayer. Conversely, TPPS4 generates singlet oxygen in the bulk whose lifetime is increased in D2O. Therefore, the affinity of the porphyrin to the membrane modulates the rate, type and degree of lipid damage.

DOI

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