Skip to main content
NCBI home page
Biochemical Journal logoLink to Biochemical Journal
. 1973 Jan;131(1):155–162. doi: 10.1042/bj1310155

The purification, composition and specificity of wheat-germ agglutinin

A K Allen 1, A Neuberger 1, N Sharon 1,*
PMCID: PMC1177449  PMID: 4737292

Abstract

1. The purification of wheat-germ agglutinin from commercial wheat germ is described. By ion-exchange chromatography three active proteins (isolectins) were separated, one of which was examined in detail. 2. The amino acid composition is unusual, as 20% of residues are half-cystine and 21% are glycine. Unlike most lectins and contrary to previous reports, this protein is not a glycoprotein. 3. The efficiency of various saccharides as inhibitors of the agglutination reaction was investigated and from this the specificity of the binding site was inferred. Of monosaccharides, only derivatives of glucose with a 2-acetamido group and a free 3-hydroxyl group are effective inhibitors, and glycosides of either anomeric configuration are bound. Oligosaccharides are much more powerful inhibitors of agglutination than are monosaccharides. 4. It is proposed that the binding site consists of three or four subsites with differing specificities, in a cleft in the molecule resembling that proposed for hen's-egg-white lysozyme.

Full text

PDF
155

Selected References

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

  1. Andrews P. The gel-filtration behaviour of proteins related to their molecular weights over a wide range. Biochem J. 1965 Sep;96(3):595–606. doi: 10.1042/bj0960595. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Blake C. C., Johnson L. N., Mair G. A., North A. C., Phillips D. C., Sarma V. R. Crystallographic studies of the activity of hen egg-white lysozyme. Proc R Soc Lond B Biol Sci. 1967 Apr 18;167(1009):378–388. doi: 10.1098/rspb.1967.0035. [DOI] [PubMed] [Google Scholar]
  3. Burger M. M., Goldberg A. R. Identification of a tumor-specific determinant on neoplastic cell surfaces. Proc Natl Acad Sci U S A. 1967 Feb;57(2):359–366. doi: 10.1073/pnas.57.2.359. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Davies R. C., Neuberger A., Wilson B. M. The dependence of lysozyme activity on pH and ionic strength. Biochim Biophys Acta. 1969 Apr 22;178(2):294–305. doi: 10.1016/0005-2744(69)90397-0. [DOI] [PubMed] [Google Scholar]
  5. Gillespie J. M., Haylett T., Lindley H. Evidence of homology in a high-sulphur protein fraction (SCMK-B2) of wool and hair alpha-keratins. Biochem J. 1968 Nov;110(2):193–200. doi: 10.1042/bj1100193. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. HIRS C. H. The oxidation of ribonuclease with performic acid. J Biol Chem. 1956 Apr;219(2):611–621. [PubMed] [Google Scholar]
  7. KAGI J. H., VALLEE B. L. Metallothionein: a cadmium and zinc-containign protein from equine renal cortex. II. Physico-chemical properties. J Biol Chem. 1961 Sep;236:2435–2442. [PubMed] [Google Scholar]
  8. LIENER I. E. The photometric determination of the hemagglutinating activity of soyin and crude soybean extracts. Arch Biochem Biophys. 1955 Jan;54(1):223–231. doi: 10.1016/0003-9861(55)90025-4. [DOI] [PubMed] [Google Scholar]
  9. Lis H., Sela B. A., Sachs L., Sharon N. Specific inhibition by N-acetyl-D-galactosamine of the interaction between soybean agglutinin and animal cell surfaces. Biochim Biophys Acta. 1970 Sep 15;211(3):582–585. doi: 10.1016/0005-2736(70)90265-8. [DOI] [PubMed] [Google Scholar]
  10. MATSUSHIMA Y., MIYAZAKI T., PARK J. T. SEPARATION OF METHYL N-ACETYL-D-GLUCOSAMINIDE ANOMERS. J Biochem. 1963 Jul;54:109–110. doi: 10.1093/oxfordjournals.jbchem.a127740. [DOI] [PubMed] [Google Scholar]
  11. Matsubara H., Sasaki R. M. High recovery of tryptophan from acid hydrolysates of proteins. Biochem Biophys Res Commun. 1969 Apr 29;35(2):175–181. doi: 10.1016/0006-291x(69)90263-0. [DOI] [PubMed] [Google Scholar]
  12. Nagata Y., Burger M. M. Wheat germ agglutinin. Isolation and crystallization. J Biol Chem. 1972 Apr 10;247(7):2248–2250. [PubMed] [Google Scholar]
  13. Ozanne B., Sambrook J. Binding of radioactively labelled concanavalin A and wheat germ agglutinin to normal and virus-transformed cells. Nat New Biol. 1971 Aug 4;232(31):156–160. doi: 10.1038/newbio232156a0. [DOI] [PubMed] [Google Scholar]
  14. Poretz R. D., Goldstein I. J. An examination of the topography of the saccharide binding sites of concanavalin A and of the forces involved in complexation. Biochemistry. 1970 Jul 7;9(14):2890–2896. doi: 10.1021/bi00816a021. [DOI] [PubMed] [Google Scholar]
  15. Reisfeld R. A., Börjeson J., Chessin L. N., Small P. A., Jr Isolation and characterization of a mitogen from pokeweek (Phytolacca americana). Proc Natl Acad Sci U S A. 1967 Nov;58(5):2020–2027. doi: 10.1073/pnas.58.5.2020. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. SPRINGER G. F., DESAI P. R., KOLECKI B. SYNTHESIS AND IMMUNOCHEMISTRY OF FUCOSE METHYL ETHERS AND THEIR METHYLGLYCOSIDES. Biochemistry. 1964 Aug;3:1076–1085. doi: 10.1021/bi00896a012. [DOI] [PubMed] [Google Scholar]
  17. Sharon N., Lis H. Lectins: cell-agglutinating and sugar-specific proteins. Science. 1972 Sep 15;177(4053):949–959. doi: 10.1126/science.177.4053.949. [DOI] [PubMed] [Google Scholar]
  18. Sharon N. The chemical structure of lysozyme substrates and their cleavage by the enzyme. Proc R Soc Lond B Biol Sci. 1967 Apr 18;167(1009):402–415. doi: 10.1098/rspb.1967.0037. [DOI] [PubMed] [Google Scholar]
  19. Shier W. T. Preparation of a "chemical vaccine" against tumor progression. Proc Natl Acad Sci U S A. 1971 Sep;68(9):2078–2082. doi: 10.1073/pnas.68.9.2078. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. So L. L., Goldstein I. J. Protein-carbohydrate interaction. 13. The interaction of concanavalin A with alpha-mannans from a variety of microorganisms. J Biol Chem. 1968 Apr 25;243(8):2003–2007. [PubMed] [Google Scholar]
  21. ZILLIKEN F., ROSE C. S., BRAUN G. A., GYORGY P. Preparation of alkyl N-acetyl-alpha and -beta-D-glucosaminides and their microbiological activity for Lactobacillus bifidus var. Penn. Arch Biochem Biophys. 1955 Feb;54(2):392–397. doi: 10.1016/0003-9861(55)90052-7. [DOI] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES