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. 1968 Feb;106(3):593–600. doi: 10.1042/bj1060593

The estimation of galactose, mannose and fucose in glycoproteins by radioisotope dilution

E R B Graham 1, A Neuberger 1
PMCID: PMC1198547  PMID: 4170711

Abstract

1. The principle of radioisotope dilution, as used previously for the estimation of mannose in egg albumin, was applied on a semi-micro scale to the estimation of fucose, mannose and galactose in some glycoproteins. The sugars were separated by partition chromatography on columns of Celite 545. 2. The release of mannose from egg albumin in 2n-hydrochloric acid at 100° after various times was determined by the radioisotope-dilution method and found to have a half-time of 7min. 3. The destruction of mannose in 2n-hydrochloric acid after 3hr. at 100° was found to be small if air was excluded. The destruction was slightly increased by the presence of lysozyme containing tryptophan in an amount equimolar with the mannose. The same amount of free tryptophan caused considerable loss of mannose. 4. Analytical values are reported for the non-amino sugar contents of egg albumin, rabbit γ-globulin and some samples of blood-group-specific substances. The values found were similar to the most reliable estimates published previously.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. CRUMPTON M. J., WILKINSON J. M. AMINO ACID COMPOSITIONS OF HUMAN AND RABBIT GAMMA-GLOBULINS AND OF THE FRAGMENTS PRODUCED BY REDUCTION. Biochem J. 1963 Aug;88:228–234. doi: 10.1042/bj0880228. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Davies J. W., Cocking E. C. Liquid scintillation counting of 14C and 3H samples using glass-fibre or filter-paper discs. Biochim Biophys Acta. 1966 Feb 28;115(2):511–513. doi: 10.1016/0304-4165(66)90456-9. [DOI] [PubMed] [Google Scholar]
  3. FLEISCHMAN J. B., PORTER R. R., PRESS E. M. THE ARRANGEMENT OF THE PEPTIDE CHAINS IN GAMMA-GLOBULIN. Biochem J. 1963 Aug;88:220–228. doi: 10.1042/bj0880220. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. FRANCOIS C., MARSHALL R. D., NEUBERGER A. Carbohydrates in protein. 4. The determination of mannose in hen's-egg albumin by radioisotope dilution. Biochem J. 1962 May;83:335–341. doi: 10.1042/bj0830335. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. GOTTSCHALK A., ADA G. L. The separation and quantitative determination of the component sugars of mucoproteins. Biochem J. 1956 Apr;62(4):681–686. doi: 10.1042/bj0620681. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Gyorky G., Houck J. C. The determination of terminal protein-bound fucose. Can J Biochem. 1965 Nov;43(11):1807–1811. doi: 10.1139/o65-201. [DOI] [PubMed] [Google Scholar]
  7. HALL R. H. Resolution of mixtures of sugars by means of partition chromatography on columns: resolution of a mixture of the four aldopentoses and 2-deoxyribose and of certain hexoses. Anal Biochem. 1962 Nov;4:395–399. doi: 10.1016/0003-2697(62)90140-9. [DOI] [PubMed] [Google Scholar]
  8. HALL R. H. Separation of nucleic acid degradation products on partition columns. J Biol Chem. 1962 Jul;237:2283–2288. [PubMed] [Google Scholar]
  9. JOHANSEN P. G., MARSHALL R. D., NEUBERGER A. Carbohydrates in protein. 2. The hexose, hexosamine, acetyl and amide-nitrogen content of hen's-egg albumin. Biochem J. 1960 Nov;77:239–247. doi: 10.1042/bj0770239. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Kekwick R. A. The serum proteins in multiple myelomatosis. Biochem J. 1940 Sep;34(8-9):1248–1257. doi: 10.1042/bj0341248. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. NOLAN C., SMITH E. L. Glycopeptides. II. Isolation and properties of glycopeptides from rabbit gamma-globulin. J Biol Chem. 1962 Feb;237:446–452. [PubMed] [Google Scholar]
  12. NUENKE R. H., CUNNINGHAM L. W. Preparation and structural studies of ovalbumin glycopeptides. J Biol Chem. 1961 Sep;236:2452–2460. [PubMed] [Google Scholar]
  13. Neuberger A. Carbohydrates in protein: The carbohydrate component of crystalline egg albumin. Biochem J. 1938 Sep;32(9):1435–1451. doi: 10.1042/bj0321435. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. PARK J. T., JOHNSON M. J. A submicrodetermination of glucose. J Biol Chem. 1949 Nov;181(1):149–151. [PubMed] [Google Scholar]
  15. PUSZTAI A., MORGAN W. T. Studies in immunochemistry. 18. The isolation and properties of a sialomucopolysaccharide possessing blood-group Le-a specificity and virus-receptor activity. Biochem J. 1961 Jan;78:135–146. doi: 10.1042/bj0780135. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Pusztai A., Morgan W. T. Studies in immunochemistry. 23. Some observations on the heterogeneity of preparations of human blood-group-specific substances. Biochem J. 1964 Nov;93(2):363–372. doi: 10.1042/bj0930363. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. SPIRO R. G. Demonstration of a single peptide chain in the glycoprotein fetuin: terminal amino acid analyses and studies of the oxidized and reduced alkylated preparations. J Biol Chem. 1963 Feb;238:644–649. [PubMed] [Google Scholar]
  18. WALBORG E. F., Jr, WARD D. N. THE CARBOHYDRATE COMPONENTS OF OVINE LUTEINIZING HORMONE. Biochim Biophys Acta. 1963 Oct 29;78:304–312. doi: 10.1016/0006-3002(63)91641-x. [DOI] [PubMed] [Google Scholar]

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