Role of band 3 tyrosine phosphorylation in the regulation of erythrocyte glycolysis.
Previous studies demonstrated that the in vitro tyrosine phosphorylation of the human erythrocyte anion transporter, band 3, prevented the binding of various glycolytic enzymes to the N terminus of the cytoplasmic tail. Since these enzymes are inhibited in their bound state, the functional consequences of band 3 tyrosine phosphorylation in the red cell should be to activate the enzymes and elevate glycolysis. We searched for various enhancers of band 3 tyrosine phosphorylation using a novel assay designed to measure the phosphotyrosine levels at the band 3 tyrosine phosphorylation/glycolytic enzyme-binding site. This assay measures the extent of phosphorylation of a synthetic band 3 peptide entrapped within resealed red cells. Using this assay, three distinct compounds, all mild oxidants, were found to stimulate the tyrosine phosphorylation of band 3. All three compounds were also found to elevate glycolytic rates in intact erythrocytes. Moreover, the antitumor drug adriamycin was found to coordinately prevent these agents from stimulating both band 3 tyrosine phosphorylation and erythrocyte glycolysis. These results suggest a possible function for a protein tyrosine kinase in human erythrocytes, to regulate glycolysis through the tyrosine phosphorylation of band 3.
Harrison
M L
ML
Department of Medicinal Chemistry and Pharmacognosy, Purdue University, West Lafayette, Indiana 47907.
Rathinavelu
P
P
Arese
P
P
Geahlen
R L
RL
Low
P S
PS
D000595:Amino Acid Sequence; D001457:Anion Exchange Protein 1, Erythrocyte / Q000378:metabolism*; D002451:Cell Compartmentation; D003958:Diamide / Q000494:pharmacology; D004317:Doxorubicin / Q000494:pharmacology; D004912:Erythrocytes / Q000378:metabolism*; D005292:Ferricyanides / Q000494:pharmacology; D006019:Glycolysis / Q000187:drug effects; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D066298:In Vitro Techniques; D007773:Lactates / Q000378:metabolism; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000737:chemistry / Q000378:metabolism; D010766:Phosphorylation; D019000:Phosphotyrosine; D014443:Tyrosine / Q000031:analogs & derivatives / Q000378:metabolism
D001457:Anion Exchange Protein 1, Erythrocyte; D005292:Ferricyanides; D007773:Lactates; D009842:Oligopeptides; D003958:Diamide; C007931:hexacyanoferrate III; D019000:Phosphotyrosine; D014443:Tyrosine; D004317:Doxorubicin; D006861:Hydrogen Peroxide