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Resealing of Protein Tyrosine Kinase Substrates into Human Erythrocytes by Rapid Freezing and Thawing in Liquid Nitrogen

  • Marietta Harrison
  • Chris Isaacson
  • Prema Rathinavelu
  • Philip S. Low
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 326)

Abstract

We have proposed a hypothesis that the rate of glycolysis in erythrocytes can be controlled in part by the phosphorylation-dependent interaction of several glycolytic enzymes with the N-terminus of the cytoplasmic domain of band 3.1-3 Briefly, as shown by Steck and coworkers4-7, glyceraldehyde-3-phosphate dehydrogenase, aldolase, and phosphofructokinase bind to an acidic sequence within the first 40 amino acids of the anion transport protein, band 3. Because the enzymes are inhibited upon association with the cytoplasmic terminus of band 3, glycolysis is retarded under conditions that promote enzyme binding. However, when the enzymes are released from their inhibitory site, substrates flow more freely down the pathway and glycolysis accelerates. Based on in vitro studies, three potentially physiological transitions can lead to displacement of the glycolytic enzymes from band 3: i) elevation of pH8,9, ii) an increase in the concentrations of certain glycolytic substrates8-10, and iii) phosphorylation of tyrosine 8 or 21 within the enzyme binding site on band 3.1,2 Because little evidence was available to support any of the above regulatory mechanisms in vivo, we sought to develop a method that would allow quantitative assessment of the phosphorylation of band 3 on tyrosine 8 in vivo.

Keywords

Human Erythrocyte Glycolytic Enzyme Peptide Solution Rapid Freezing Enzyme Binding 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Springer Science+Business Media New York 1992

Authors and Affiliations

  • Marietta Harrison
    • 1
  • Chris Isaacson
    • 1
  • Prema Rathinavelu
    • 2
  • Philip S. Low
    • 2
  1. 1.Department of Medicinal Chemistry and PharmacognosyUSA
  2. 2.Department of ChemistryPurdue UniversityIndianaUSA

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