PH-dependence of detergent-induced hemolysis and vesiculation of erythrocytes.
The influence of pH of the medium on the parameters of detergent-induced fast hemolysis and vesiculation of human erythrocytes was studied. In the range of pH 6.3-7.2 neither the extent nor the rate of the vesiculation induced by 25 microM sodium dodecyl sulfate (SDS) changed. However, a decrease of pH from 8.0 to 5.8 strongly modified both the extent and the rate of the hemolysis induced by SDS. Within the range of pH 8.0-6.4, the effect can be ascribed to the increase of the positive charge of the membrane. This could lead to the accumulation of the membrane-bound anion detergent and, hence, to the change of the hemolysis parameters. Non-charged detergent Triton X-100 did not display any pH-dependence. At pH between 6.4 and 5.8 the extent and rate of hemolysis changed in a complicated manner. The kinetic curves of hemolysis could be approximated by a single exponential within the pH range between 8.0 and 7.2. Upon further reduction of pH, a second exponential component, with a larger time constant, appeared in the kinetic curves. At 5.8 < pH < 7.2, the contribution of the "fast" hemolysis dropped virtually to zero, with pK about 6.0. This points to a structural transition of the membrane, possibly involving histidine. We suggest that the parameters of the detergent-induced hemolysis are sensitive to the changes of the charge and structural state of erythrocyte membrane.
Chernitsky
E A
EA
Institute of Photobiology, National Academy of Sciences of Belarus, Minsk, Republic of Belarus. sei@biobel.bas-net.by
Rozin
V V
VV
Senkovich
O A
OA
D000110:Acetylcholinesterase / Q000378:metabolism; D003902:Detergents / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004912:Erythrocytes / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure*; D006461:Hemolysis; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008433:Mathematics; D017830:Octoxynol / Q000494:pharmacology*; D010758:Phosphorus / Q000378:metabolism; D012967:Sodium Dodecyl Sulfate / Q000494:pharmacology*; D013053:Spectrophotometry; D013501:Surface-Active Agents / Q000494:pharmacology
D003902:Detergents; D013501:Surface-Active Agents; D010758:Phosphorus; D012967:Sodium Dodecyl Sulfate; D017830:Octoxynol; D000110:Acetylcholinesterase