Effects of temperature and pH on hemoglobin release from hydrostatic pressure-treated erythrocytes.
The release of hemoglobin from human erythrocytes hemolyzed beforehand by hydrostatic pressure, osmotic pressure, and freeze-thaw methods was examined as a function of temperature (0-45 degrees C) and pH (5.5-8.8) at atmospheric pressure. Only in the case of high pressure (2,000 bar) did the release of hemoglobin increase significantly with decreasing temperature and pH. Maleimide spin label studies showed that the temperature and pH dependences of hemoglobin release were qualitatively explicable in terms of those of the conformational changes of membrane proteins. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of membrane proteins showed the diminution of band intensities corresponding to spectrin, ankyrin, and actin in the erythrocytes hemolyzed by high pressure. Cross-linking of cytoskeletal proteins by diamide stabilized the membrane structure against high pressure and suppressed hemoglobin release. These results indicate that the disruption of cytoskeletal apparatus by high pressure makes the membrane more leaky.
Yamaguchi
T
T
Department of Chemistry, Faculty of Science, Fukuoka University.
Kawamura
H
H
Kimoto
E
E
Tanaka
M
M
D003432:Cross-Linking Reagents / Q000378:metabolism; D003599:Cytoskeleton / Q000378:metabolism; D003958:Diamide / Q000378:metabolism; D004578:Electron Spin Resonance Spectroscopy; D004591:Electrophoresis, Polyacrylamide Gel; D004910:Erythrocyte Membrane / Q000378:metabolism; D004912:Erythrocytes / Q000378:metabolism*; D005615:Freezing; D006454:Hemoglobins / Q000378:metabolism*; D006461:Hemolysis / Q000502:physiology; D006801:Humans; D006863:Hydrogen-Ion Concentration; D006874:Hydrostatic Pressure; D008301:Maleimides / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism; D009997:Osmotic Pressure; D011312:Pressure; D013113:Spin Labels; D013696:Temperature
D003432:Cross-Linking Reagents; D006454:Hemoglobins; D008301:Maleimides; D008565:Membrane Proteins; D013113:Spin Labels; D003958:Diamide; C043592:maleimide