title: Effect of oxidative stress and erythropoietin on cytoskeletal protein and lipid organization in human erythrocytes.
abstract: Phenylhydrazine (PHX)-mediated damage in human red blood cells has been assessed by monitoring the release of tyrosine from cell proteins as well as using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). PHX-treated red blood cells exhibited concentration- and time-dependent tyrosine release. ATP has no effect on the release of tyrosine. This observation is supported by SDS-PAGE pattern of RBC membrane proteins, which shows a correlation between tyrosine release and cytoskeletal protein degradation. PHX requires the presence of erythrocyte cytosolic fraction for the degradation, possibly due to the presence of a proteolytic enzyme in the cytosol. PHX treatment renders the membrane proteins susceptible to the proteolytic attack. Treatment of PHX-exposed erythrocyte with bee venom phospholipase A2 induces the translocation of phosphatidylserine (PS) and phosphatidylethanolamine (PE) to the outer surface of the cell membrane. At the same time, phosphatidylcholine (PC) was translocated towards the inner surface, altering the membrane phospholipid asymmetry. Interestingly, increased tyrosine production followed by translocation of phospholipids across the red blood cell membrane by PHX treatment is completely inhibited by 0.2 units of erythropoietin (EP). Our findings suggest that exposure of red blood cells to an oxidant like PHX causes degradation of cytoskeletal protein by an ATP-independent proteolytic pathway and this in turn allows the transbilayer movement of phospholipids across the cell membrane. EP, by scavenging the hydroxyl radicals produced during interaction of PHX with red blood cells, protects the erythrocytes from oxidative attack.
authors.affiliation: Department of Life Science and Biotechnology, Jadavpur University, Calcutta, India.
authors.forename: T D
authors.initials: TD
authors.lastname: Choudhury
authors.forename: N
authors.initials: N
authors.lastname: Das
authors.forename: A
authors.initials: A
authors.lastname: Chattopadhyay
authors.forename: A G
authors.initials: AG
authors.lastname: Datta
mesh_terms: D003598:Cytoskeletal Proteins / Q000097:blood*; D004228:Dithionitrobenzoic Acid / Q000494:pharmacology; D004910:Erythrocyte Membrane / Q000187:drug effects / Q000378:metabolism; D004912:Erythrocytes / Q000187:drug effects* / Q000378:metabolism*; D004921:Erythropoietin / Q000097:blood*; D005033:Ethylmaleimide / Q000494:pharmacology; D006801:Humans; D015227:Lipid Peroxidation / Q000187:drug effects; D008563:Membrane Lipids / Q000097:blood*; D018384:Oxidative Stress; D010659:Phenylhydrazines / Q000494:pharmacology; D013439:Sulfhydryl Reagents / Q000494:pharmacology; D014443:Tyrosine / Q000097:blood; D020245:p-Chloromercuribenzoic Acid / Q000494:pharmacology
chemical_list: D003598:Cytoskeletal Proteins; D008563:Membrane Lipids; D010659:Phenylhydrazines; D013439:Sulfhydryl Reagents; C030299:phenylhydrazine; D004921:Erythropoietin; D014443:Tyrosine; D020245:p-Chloromercuribenzoic Acid; D004228:Dithionitrobenzoic Acid; D005033:Ethylmaleimide