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Nuclear transport of photosensitizers during photosensitization and oxidative stress
Author(s) -
Patito Ifat Amit,
Rothmann Chana,
Malik Zvi
Publication year - 2001
Publication title -
biology of the cell
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.543
H-Index - 85
eISSN - 1768-322X
pISSN - 0248-4900
DOI - 10.1016/s0248-4900(01)01118-2
Subject(s) - photosensitizer , sephadex , porphyrin , fluorescence , oxidative stress , biology , nucleolus , flow cytometry , nucleus , biophysics , fluorescence microscope , biochemistry , microbiology and biotechnology , chemistry , photochemistry , quantum mechanics , enzyme , physics
Summry— The nuclear transport pathways of the photosensitizers meso ‐tetra(4‐sulfonatophenyl)porphyrin (TPPS 4 ) and meso ‐tetra(4‐N‐methylpyridyl)porphyrin (TMPyP) during photosensitization and oxidative stress were characterized in CT‐26 murine colon carcinoma cells using fluorescence microscopy and multi‐pixel spectral imaging. Prior to irradiation, TPPS 4 and TMPyP localized mainly in the lysosomes, while irradiation or H 2 O 2 treatment induced a relocalization into the nucleus and nucleoli. Flow cytometry analysis of isolated nuclei from the treated cells showed an increase in nuclear fluorescence accompanying the relocalization. Isolation and separation of the nuclear proteins according to molecular weight was performed using a sephadex G‐100 column. The protein fractions exhibiting high fluorescence were separated by high performance liquid chromatography. Five major classes of proteins with a retention time of 1, 7, 11, 12 and 15 min were obtained. Each photosensitizer was associated with a distinct class of proteins. While TPPS 4 fluorescence was detected in the protein fraction with a retention time of 11 min, TMPyP fluorescence was associated with a protein fraction having a retention time of 7 min. We conclude that although oxidative stress triggers entry into the nucleus of both TPPS 4 and TMPyP, each sensitizer uses a distinct transport mechanism based on its chemical properties.