title: Subcellular fractionation of stored red blood cells reveals a compartment-based protein carbonylation evolution.
abstract: During blood banking, erythrocytes undergo storage lesions, altering or degrading their metabolism, rheological properties, and protein content. Carbonylation is a hallmark of protein oxidative lesions, thus of red blood cell oxidative stress. In order to improve global erythrocyte protein carbonylation assessment, subcellular fractionation has been established, allowing us to work on four different protein populations, namely soluble hemoglobin, hemoglobin-depleted soluble fraction, integral membrane and cytoskeleton membrane protein fractions. Carbonylation in erythrocyte-derived microparticles has also been investigated. Carbonylated proteins were derivatized with 2,4-dinitrophenylhydrazine (2,4-DNPH) and quantified by western blot analyses. In particular, carbonylation in the cytoskeletal membrane fraction increased remarkably between day 29 and day 43 (P<0.01). Moreover, protein carbonylation within microparticles released during storage showed a two-fold increase along the storage period (P<0.01). As a result, carbonylation of cytoplasmic and membrane protein fractions differs along storage, and the present study allows explaining two distinct steps in global erythrocyte protein carbonylation evolution during blood banking. This article is part of a Special Issue entitled: Integrated omics.
authors.affiliation: Service Régional Vaudois de Transfusion Sanguine, route de Corniche 2, CH-1066 Epalinges, Switzerland.
authors.forename: Julien
authors.initials: J
authors.lastname: Delobel
authors.forename: Michel
authors.initials: M
authors.lastname: Prudent
authors.forename: Olivier
authors.initials: O
authors.lastname: Rubin
authors.forename: David
authors.initials: D
authors.lastname: Crettaz
authors.forename: Jean-Daniel
authors.initials: JD
authors.lastname: Tissot
authors.forename: Niels
authors.initials: N
authors.lastname: Lion
mesh_terms: D001793:Blood Preservation; D055252:Cell-Derived Microparticles / Q000378:metabolism*; D004912:Erythrocytes / Q000166:cytology / Q000378:metabolism*; D006801:Humans; D050050:Protein Carbonylation; D013997:Time Factors