Behavior of O-Glycosyl and O-Phosphoryl Proteins in Alkaline Solution

  • John R. Whitaker
  • Robert E. Feeney
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 86)


0-Glycosyl and 0-phosphoryl groups, as well as disulfide bonds, are rapidly removed from proteins in alkaline solution primarily via β-elimination. The reaction is initiated by abstraction of the α-hydrogen of an amino acid residue by hydroxide ion. The carbanion undergoes rearrangement expelling the glyco- or phosphoryl group resulting in formation of a dehydroalanyl (from serine) or β-methyldehy-droalanyl (from threonine) residue. The unsaturated derivatives are reactive with internal protein nucleophilic groups and with external nucleophiles. These addition reactions, some leading to crosslinking, result in changed properties of the protein. Several factors may affect rates of β-elimination and addition. For these studies, we used two unique proteins: phosvitin, a well-characterized protein with 120 0-phosphoryl groups and no cystine or 0-glycosyl groups; and a glycopeptide related to the antifreeze protein from Antarctic fish, a well-characterized protein consisting of (Ala-Ala-Thr)n in which all of the threonyl residues are glycosylated. At the dilute concentrations of proteins used, rates of β-elimination and addition were independent of protein concentration but directly dependent upon the hydroxide ion concentration. With phosvitin, rates of β-elimination and addition were quite dependent on ionic strength and the rate of β-elimination was increased 20-fold in the presence of Ca2+. Activation energies for both β-elimination and addition were near 20 kcal/mole. Implications of these reactions for protein chemistry and protein processing will be discussed.


Carbohydrate Cysteine Serine Lysine Arginine 
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Copyright information

© Springer Science+Business Media New York 1977

Authors and Affiliations

  • John R. Whitaker
    • 1
  • Robert E. Feeney
    • 1
  1. 1.Department of Food Science and TechnologyUniversity of CaliforniaDavisUSA

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