The influence of cupric ions on porphyrin-induced photodynamic membrane damage in human red blood cells.
Photooxidation of various susceptible substrates with hematoporphyrin derivative (photofrin) as sensitizer was strongly inhibited by simultaneous addition of cupric acetate to the reaction mixture. With sulfhydryl-containing compounds, however, an increased rate of photooxidation was observed under these experimental conditions. Preincubation of photofrin and cupric acetate at equimolar concentrations for 24 h at room temperature yielded a stable photofrin-Cu2+ complex. This complex did not act as photosensitizer with histidine, tryptophan, tyrosine, methionine or guanosine as substrates. With dithiothreitol, however, the photofrin-Cu2+ complex still acted as a photosensitizer, with an efficiency of about 30% as compared to free photofrin. Also in red blood cell membranes only sulfhydryl groups were photooxidized with the photofrin-Cu2+ complex as sensitizer. Illumination of intact erythrocytes in the presence of the photofrin-Cu2+ complex resulted in K+ leakage and, ultimately, photohemolysis. Pretreatment of the cells with N-ethylmaleimide and 5,5'-dithiobis(2-nitrobenzoic acid) (DTNB) inhibited this photodynamically induced K+ loss. Considering recent studies on the reactivity of distinct membrane SH-groups with various sulfhydryl reagents this suggests that a sulfhydryl group, located in the 17 kDa membrane-bound fragment of band 3, is involved in photodynamic K+ leakage with the photofrin-Cu2+ complex as sensitizer.
van Steveninck
J
J
Boegheim
J P
JP
Dubbelman
T M
TM
D003300:Copper / Q000494:pharmacology*; D004228:Dithionitrobenzoic Acid / Q000494:pharmacology; D004910:Erythrocyte Membrane / Q000187:drug effects* / Q000378:metabolism / Q000648:ultrastructure; D005033:Ethylmaleimide / Q000494:pharmacology; D017324:Hematoporphyrin Derivative; D006415:Hematoporphyrins / Q000494:pharmacology*; D006801:Humans; D007700:Kinetics; D009942:Organometallic Compounds; D010777:Photochemistry; D011188:Potassium / Q000097:blood
D006415:Hematoporphyrins; D009942:Organometallic Compounds; C015092:cupric acetate; D017324:Hematoporphyrin Derivative; D003300:Copper; D004228:Dithionitrobenzoic Acid; D005033:Ethylmaleimide; D011188:Potassium