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Reduction by Mercaptoethanol and Proteolysis of the Non-Glycosylated Peptide Region of Pig Gastric Mucus Glycoprotein

  • Jeffrey P. Pearson
  • Adrian Allen
Chapter
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 144)

Abstract

The native pig gastric mucus glycoprotein was purified free from non-covalently bound protein by a combination of gel-filtration on Sepharose 4B and equilibrium centrifugation in a CsCl gradient (Starkey et al., 1974; Pearson et al., 1981). On reduction with 0.2M mercaptoethanol the native glycoprotein (2 × 106 molecular weight) is split into an average of 4 equal sized subunits (5 × 105 molecular weight) and a 70,000 molecular weight protein fraction. The 70,000 molecular weight protein fraction which contains little or no carbohydrate can be separated from the reduced glycoprotein subunits by equilibrium centrifugation in a CsCl gradient (Allen et al., 1980; Pearson & Allen, 1980). Here we report further investigations on the structure of the non-glycosylated region of glycoprotein defined as that part of the peptide core which is free of carbohydrate and is removed when the molecule is split by proteolysis into 4 subunits of molecular weight 5 × 105 (Scawen & Allen, 1977).

Keywords

Cystic Fibrosis Amino Acid Analysis Amino Acid Content Molecular Weight Protein Peptide Core 
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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References

  1. Allen, A., Mantle, M. and Pearson, J.P., 1980, The structure and properties of mucus glycoproteins. In: Perspectives in Cystic Fibrosis, edited by J. Sturgess, pp. 102–112. Canadian Cystic Fibrosis Foundation, Toronto.Google Scholar
  2. Pearson, J.P., Allen, A. and Venables, C.W., 1980, Gastric mucus: isolation and polymeric structure of the undegraded glycoprotein: its breakdown by pepsin. Gastroenterology, 78: 709–715.PubMedGoogle Scholar
  3. Pearson, J.P., Allen, A. and Parry, S., 1981, A protein 70,000 molecular weight, isolated from purified pig gastric mucus glycoprotein by reduction of disulphide bridges: implication in polymeric structure. Biochem. J. in press.Google Scholar
  4. Scawen, M. and Allen, A., 1977, The action of proteolytic enzymes on the glycoprotein from pig gastric mucus. Biochem. J. 163: 363–368.PubMedGoogle Scholar
  5. Starkey, B.J., Snary, B. and Allen, A., 1974, Characterisation of gastric mucoproteins isolated by equilibrium density gradient centrifugation in CsCl. Biochem. J., 141: 633–639.PubMedGoogle Scholar

Copyright information

© Plenum Press, New York 1982

Authors and Affiliations

  • Jeffrey P. Pearson
    • 1
  • Adrian Allen
    • 1
  1. 1.Department of PhysiologyThe UniversityNewcastle upon TyneUK

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