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Lysis of resealed erythrocyte ghosts by photoactivated tetrapyrroles: Estimation of photolesion dimensions
Affiliation:1. WildEarth Guardians, PO Box 7516, Missoula, MT 59807, United States;2. Human-Environment Systems, Boise State University, 1910 University Drive, Boise, ID 83725, United States;3. Research Unit of Biodiversity (UO/CSIC/PA), Oviedo University, 33600 Mieres, Spain;4. Norwegian Institute for Nature Research, PO Box 5685 Torgard, NO-7485 Trondheim, Norway
Abstract:
  • 1.1. Resealed erythrocyte ghosts containing Na+ and glucose-6-P (G6P) as markers of membrane integrity were used as a model system for probing the damaging effects of photoactivated tetrapyrroles on cell membranes.
  • 2.2. Continuous blue-light irradiation of bilirubin (BR)-sensitized and protoporphyrin (PP)-sensitized ghosts made them progressively more permeable to Na+ the cation emerging well ahead of G6P.
  • 3.3. G6P efflux occurred abruptly after a lag period and resembled an all-or-none process.
  • 4.4. These and other results suggest that a relatively subtle structural modification (possibly in some crucial protein(s) is sufficient for Na+ release, whereas gross disruption of the bilayer (probably by free-radical lipid peroxidation) is necessary for G6P release.
  • 5.5. The dimensions of the G6P-releasing photolesions were estimated by density floatation centrifugation, using saccharides of increasing molecular size. Both BR and PP produced pores > 11 Å but <42 Å in diam, which is considerably smaller than the size range estimated in hypotonically lysed ghosts.
Keywords:
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