title: Band 3 is a target protein of spectrin's E2/E3 activity: implication for sickle cell disease and normal red blood cell aging.
abstract: We have demonstrated that a 125 kDa red blood cell (RBC) membrane protein, being a target of spectrin's E2/E3 activity, is ubiquitinated band 3. This demonstration was based on copurification of this biotinylated-ubiquitinated protein with band 3, immunoprecipitation with band 3 antibody and analysis of proteins associated with strepavidin sepharose by micro liquid chromatography coupled to tandem mass spectrometry (microLC/MS/MS). Further, we demonstrated the presence of ubiquitinated band 3 in vivo by Western blotting of purified band 3 with a monoclonal antibody (FK2) against ubiquitin. The implications of these results for sickle cell disease and RBC aging are discussed.
authors.affiliation: Department of Molecular and Cell Biology, UTD Sickle Cell Disease Research Center, Institute of Biomedical Sciences and Technology, The University of Texas at Dallas, Richardson, TX 75083, USA.
authors.forename: T L
authors.initials: TL
authors.lastname: Chang
authors.forename: F F
authors.initials: FF
authors.lastname: Cubillos
authors.forename: D G
authors.initials: DG
authors.lastname: Kakhniashvili
authors.forename: S R
authors.initials: SR
authors.lastname: Goodman
mesh_terms: D000755:Anemia, Sickle Cell / Q000097:blood / Q000378:metabolism*; D001457:Anion Exchange Protein 1, Erythrocyte / Q000032:analysis / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D004905:Erythrocyte Aging; D004910:Erythrocyte Membrane / Q000378:metabolism; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods; D008970:Molecular Weight; D013049:Spectrin / Q000032:analysis / Q000378:metabolism*; D014452:Ubiquitins / Q000378:metabolism
chemical_list: D001457:Anion Exchange Protein 1, Erythrocyte; D014452:Ubiquitins; D013049:Spectrin