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. 1979 Nov 1;183(2):213–221. doi: 10.1042/bj1830213

Lectin-binding proteins in central-nervous-system myelin. Binding of glycoproteins in purified myelin to immobilized lectins

Richard H Quarles 1, Laurence J McIntyre 1, Carol F Pasnak 1
PMCID: PMC1161549  PMID: 534494

Abstract

The capacities of immature and mature rat brain myelin, bovine myelin and human myelin to be agglutinated by soya-bean agglutinin, Ricinus communis agglutinin, wheatgerm agglutinin, and Lotus tetragonolobus agglutinin were examined. The first two lectins, which are specific for galactose and N-acetylgalactosamine, strongly agglutinated immature and mature rat myelin, weakly agglutinated bovine myelin, but did not affect human myelin. The other myelin and lectin combinations resulted in very weak or no agglutination. [3H]Fucose-labelled glycoproteins of purified adult rat brain myelin were solubilized with sodium dodecyl sulphate and allowed to bind to concanavalin A–Sepharose and each of the other lectins mentioned above, which had been immobilized on agarose. About 60% of the radioactive fucose was in glycoproteins that bound to concanavalin A–Sepharose and these glycoproteins could be eluted with solutions containing methyl α-d-mannoside and sodium dodecyl sulphate. Periodate/Schiff staining or radioactive counting of analytical gels showed that most of the major myelin-associated glycoprotein (apparent mol.wt. approx. 100000) bound to the concanavalin A, whereas the glycoproteins that did not bind were mostly of lower molecular weight. Preparative polyacrylamide-gel electrophoresis of the glycoprotein fraction that was eluted with methyl α-d-mannoside yielded a relatively pure preparation of the myelin-associated glycoprotein. Similar results were obtained with each of the other lectins, i.e. the myelin-associated glycoprotein was in the fraction that bound to the immobilized lectin. Double-labelling experiments utilizing [3H]fucose-labelled glycoproteins from adult myelin and [14C]fucose-labelled glycoproteins from 14-day-old rat brain myelin did not reveal any difference in the binding of the mature and immature glycoproteins to any of the immobilized lectins. The results in this and the preceding paper [McIntyre, Quarles & Brady (1979) Biochem. J. 183, 205–212] suggest that the myelin-associated glycoprotein is one of the principal receptors for concanavalin A and other lectins in myelin, and that this property can be utilized for the purification of this glycoprotein.

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Selected References

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  1. Eisenbarth G. S., Ruffolo R. R., Jr, Walsh F. S., Nirenberg M. Lactose sensitive lectin of chick retina and spinal cord. Biochem Biophys Res Commun. 1978 Aug 29;83(4):1246–1252. doi: 10.1016/0006-291x(78)91355-4. [DOI] [PubMed] [Google Scholar]
  2. Greenfield S., Norton W. T., Morell P. Quaking mouse: isolation and characterization of myelin protein. J Neurochem. 1971 Nov;18(11):2119–2128. doi: 10.1111/j.1471-4159.1971.tb05070.x. [DOI] [PubMed] [Google Scholar]
  3. Matthieu J. M., Brady R. O., Quarles R. H. Anomalies of myelin-associated glycoproteins in 'Quaking' mice. J Neurochem. 1974 Feb;22(2):291–296. doi: 10.1111/j.1471-4159.1974.tb11592.x. [DOI] [PubMed] [Google Scholar]
  4. Matthieu J. M., Brady R. O., Quarles R. H. Change in a myelin-associated glycoprotein in rat brain during development: metabolic aspects. Brain Res. 1975 Mar 14;86(1):55–65. doi: 10.1016/0006-8993(75)90637-x. [DOI] [PubMed] [Google Scholar]
  5. Matthieu J. M., Daniel A., Quarles R. H., Brady R. O. Interactions of concanavalin A and other lectins with CNS myelin. Brain Res. 1974 Dec 6;81(2):348–353. doi: 10.1016/0006-8993(74)90951-2. [DOI] [PubMed] [Google Scholar]
  6. Matthieu J. M., Reier P. J., Sawchak J. A. Proteins of rat brain myelin in neonatal hypothyroidism. Brain Res. 1975 Feb 14;84(3):443–451. doi: 10.1016/0006-8993(75)90764-7. [DOI] [PubMed] [Google Scholar]
  7. McIntyre L. J., Quarles R. H., Brady R. O. Lectin-binding proteins in central-nervous-system myelin. Detection of glycoproteins of purified myelin on polyacrylamide gels by [3h]concanavalin A binding. Biochem J. 1979 Nov 1;183(2):205–212. doi: 10.1042/bj1830205. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. McIntyre L. J., Quarles R. H., Brady R. O. Regional studies of myelin-associated glycoprotein in the rat central nervous system. Brain Res. 1978 Jun 23;149(1):251–256. doi: 10.1016/0006-8993(78)90607-8. [DOI] [PubMed] [Google Scholar]
  9. Nicolson G. L. The interactions of lectins with animal cell surfaces. Int Rev Cytol. 1974;39:89–190. doi: 10.1016/s0074-7696(08)60939-0. [DOI] [PubMed] [Google Scholar]
  10. Quarles R. H. Effects of pronase and neuraminidase treatment on a myelin-associated glycoprotein in developing brain. Biochem J. 1976 Apr 15;156(1):143–150. doi: 10.1042/bj1560143. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Quarles R. H., Everly J. L., Brady R. O. Evidence for the close association of a glycoprotein with myelin in rat brain. J Neurochem. 1973 Nov;21(5):1177–1191. doi: 10.1111/j.1471-4159.1973.tb07573.x. [DOI] [PubMed] [Google Scholar]
  12. Quarles R. H., Everly J. L., Brady R. O. Myelin-associated glycoprotein: a developmental change. Brain Res. 1973 Aug 30;58(2):506–509. doi: 10.1016/0006-8993(73)90022-x. [DOI] [PubMed] [Google Scholar]
  13. 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]
  14. Simpson D. L., Thorne D. R., Loh H. H. Developmentally regulated lectin in neonatal rat brain. Nature. 1977 Mar 24;266(5600):367–369. doi: 10.1038/266367a0. [DOI] [PubMed] [Google Scholar]
  15. Sternberger N. H., Quarles R. H., Itoyama Y., Webster H. D. Myelin-associated glycoprotein demonstrated immunocytochemically in myelin and myelin-forming cells of developing rat. Proc Natl Acad Sci U S A. 1979 Mar;76(3):1510–1514. doi: 10.1073/pnas.76.3.1510. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Zanetta J. P., Sarlieve L. L., Mandel P., Vincendon G., Gombos G. Fractionation of glycoproteins associated to adult rat brain myelin fractions. J Neurochem. 1977 Nov;29(5):827–838. doi: 10.1111/j.1471-4159.1977.tb10725.x. [DOI] [PubMed] [Google Scholar]
  17. Zimmerman A. W., Matthieu J. M., Quarles R. H., Brady R. O., Hsu J. M. Hypomyelination in copper-deficient rats. Prenatal and postnatal copper replacement. Arch Neurol. 1976 Feb;33(2):111–119. doi: 10.1001/archneur.1976.00500020039007. [DOI] [PubMed] [Google Scholar]

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