title: Damage to human erythrocytes by radiation-generated HO* radicals: molecular changes in erythrocyte membranes.
abstract: The effectiveness of radiation-generated HO* radicals in initiating erythrocyte hemolysis in the presence of oxygen and under anaerobic conditions and prehemolytic structural changes in the plasma-erythrocyte membrane were studied. Under anaerobic conditions the efficacy of HO* radicals in induction of hemolysis was 16-fold lower than under air. In both conditions, hemolysis was the final consequence of changes of the erythrocyte membrane. Preceding hemolysis, the dominating process under anaerobic conditions was the aggregation of membrane proteins. The aggregates were principally formed by -S-S- bridges. A decrease in spectrin and protein of band 3 content suggests their participation in the formation of the aggregates. These processes were accompanied by changes in protein conformation determined by means of 4-maleimido-2,2,6,6-tetramethylpiperidine-N-oxyl (MSL) spin label attached to membrane proteins. Under anaerobic conditions, in the range of prehemolytical doses, the reaction of HO* with lipids caused a slight (10-16%) increase in fluidity of the lipid bilayer in its hydrophobic region with a lack of lipid peroxidation. However, in the presence of oxygen, hemolysis was preceded by intense lipid peroxidation and by profound changes in the conformation of membrane proteins. At the radiation dose that normally initiates hemolysis a slight aggregation of proteins was observed. Changes were not observed in particular protein fractions. It can be suggested the cross-linking induced by HO* radicals under anaerobic conditions and a lack of lipid peroxidation are the cause of a decrease in erythrocyte sensitivity to hemolysis. Contrary, under aerobic conditions, molecular oxygen suppresses cross-linking, catalysing further steps of protein and lipid oxidation, which accelerate hemolysis.
authors.affiliation: Department of Molecular Biophysics, University of Lodz, Banacha 12/16, 90-237 Lodz, Poland.
authors.forename: Zofia
authors.initials: Z
authors.lastname: Szweda-Lewandowska
authors.forename: Anita
authors.initials: A
authors.lastname: Krokosz
authors.forename: Marta
authors.initials: M
authors.lastname: Gonciarz
authors.forename: Wieslawa
authors.initials: W
authors.lastname: Zajeczkowska
authors.forename: Mieczysław
authors.initials: M
authors.lastname: Puchała
mesh_terms: D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D003497:Cyclic N-Oxides / Q000494:pharmacology; D004249:DNA Damage; D003720:Densitometry; D004307:Dose-Response Relationship, Radiation; D004591:Electrophoresis, Polyacrylamide Gel; D004910:Erythrocyte Membrane / Q000378:metabolism; D004912:Erythrocytes / Q000166:cytology* / Q000378:metabolism / Q000473:pathology*; D006461:Hemolysis; D006801:Humans; D017665:Hydroxyl Radical; D008051:Lipid Bilayers; D050356:Lipid Metabolism; D015227:Lipid Peroxidation; D008055:Lipids / Q000737:chemistry; D010100:Oxygen / Q000737:chemistry / Q000378:metabolism; D011487:Protein Conformation; D017382:Reactive Oxygen Species; D013113:Spin Labels
chemical_list: D003497:Cyclic N-Oxides; D008051:Lipid Bilayers; D008055:Lipids; D017382:Reactive Oxygen Species; D013113:Spin Labels; C017480:4-maleimido-2,2,6,6-tetramethylpiperidinooxyl; D017665:Hydroxyl Radical; D010100:Oxygen