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Published on: November 5, 2019
Band 3 is a target protein of spectrin's E2/E3 activity: implication for sickle cell disease and normal red blood
T L Chang1, F F Cubillos, D G Kakhniashvili
1Department 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.
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.
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