title: Oxidative stress modulates the organization of erythrocyte membrane cytoskeleton.
abstract: BACKGROUND
  Apart from their main role in transporting oxygen and carbon dioxide, erythrocytes play also an important role in organism antioxidative defence. Direct exposure to reactive oxygen species (ROS) results in shortening of their half-life, even by 50%. The presence of glucose, being the substrate in pentose phosphate pathway (PPP) cycle, is one of the factors that can have influence on the level of oxidative stress. The activity of PPP increases during oxidative stress. Glucose guarantees normal PPP functioning with the production of reductive equivalents in the amounts necessary to reproduction of glutathione--nonenzymatic free radical scavenger. In available literature there are no reports regarding the changes in protein contents of erythrocyte cytoskeleton exposed to t-butyl hydroperoxide in relation to glucose presence in incubation medium.
  
  
  METHODS
  Erythrocytes taken from 10 healthy subjects were used to assess the influence of generated free radicals on erythrocyte proteins and chosen parameters of oxidative stress. Erythrocytes were incubated in the solutions containing deferent concentrations of t-butyl hydroperoxide and glucose. Electrophoresis was performed on polyacrylamide gel in denaturating conditions. The contents of tryptophan in membranes was evaluated spectrofluorometrically.
  
  
  CONCLUSIONS
  In vitro conditions oxidative stress leads to protein damage in erythrocyte cytoskeleton, both in proteins inside the cell as well as having contact with extracellular environment. In consequence, the amount of low-molecular proteins--mainly globin, which bind to cytoskeleton, increases. This process takes place independently of glucose presence in incubation medium. One of the element of protein cytoskeleton, tryptophan, also undergoes degradation. The decrease of its contents is higher during erythrocyte exposure to t-BOOH in environment containing glucose, what can suggest prooxidative influence of glucose in conditions in vitro.
authors.affiliation: Department of Medical Chemistry, Pomeranian Medical University, Szczecin, Poland. maryla.olszewska@wp.pl
authors.forename: Maria
authors.initials: M
authors.lastname: Olszewska
authors.forename: Jerzy
authors.initials: J
authors.lastname: Wiatrow
authors.forename: Joanna
authors.initials: J
authors.lastname: Bober
authors.forename: Ewa
authors.initials: E
authors.lastname: Stachowska
authors.forename: Edyta
authors.initials: E
authors.lastname: Gołembiewska
authors.forename: Katarzyna
authors.initials: K
authors.lastname: Jakubowska
authors.forename: Małgorzata
authors.initials: M
authors.lastname: Stańczyk-Dunaj
authors.forename: Maria
authors.initials: M
authors.lastname: Pietrzak-Nowacka
mesh_terms: D001798:Blood Proteins / Q000737:chemistry; D003599:Cytoskeleton / Q000378:metabolism*; D004910:Erythrocyte Membrane / Q000737:chemistry* / Q000378:metabolism*; D016166:Free Radical Scavengers / Q000097:blood; D005609:Free Radicals / Q000378:metabolism; D005947:Glucose / Q000378:metabolism*; D005978:Glutathione / Q000378:metabolism; D006207:Half-Life; D006801:Humans; D066298:In Vitro Techniques; D018384:Oxidative Stress; D010427:Pentose Phosphate Pathway; D017382:Reactive Oxygen Species / Q000378:metabolism; D012016:Reference Values; D014364:Tryptophan / Q000737:chemistry; D020122:tert-Butylhydroperoxide / Q000378:metabolism
chemical_list: D001798:Blood Proteins; D016166:Free Radical Scavengers; D005609:Free Radicals; D017382:Reactive Oxygen Species; D014364:Tryptophan; D020122:tert-Butylhydroperoxide; D005978:Glutathione; D005947:Glucose