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. 1992 Jun 1;284(Pt 2):377–380. doi: 10.1042/bj2840377

The antigenicity of the carbohydrate moiety of an insect glycoprotein, honey-bee (Apis mellifera) venom phospholipase A2. The role of alpha 1,3-fucosylation of the asparagine-bound N-acetylglucosamine.

C Prenner 1, L Mach 1, J Glössl 1, L März 1
PMCID: PMC1132648  PMID: 1376112

Abstract

A rabbit polyclonal antiserum raised against honey-bee (Apis mellifera) venom phospholipase A2 (PLA2) contains antibodies that react exclusively with its glycosylated variants and cross-react with plant glycoproteins. The interaction of anti-(horseradish peroxidase) antiserum with PLA2 suggests the existence of a carbohydrate determinant common to both glycoproteins. E.l.i.s.a. binding and inhibition experiments, employing glycoproteins and glycopeptides of plant and animal origin with known N-glycan structures, in combination with chemical and enzymic deglycosylation, identified alpha 1,3-fucosylation of the asparagine-bound N-acetylglucosamine as the antigenic determinant. This fucose residue is present in the N-glycan of PLA2 and is frequently found in plant glycoproteins, whereas mammalian glycoproteins lack this modification.

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Selected References

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  1. Ashford D., Dwek R. A., Welply J. K., Amatayakul S., Homans S. W., Lis H., Taylor G. N., Sharon N., Rademacher T. W. The beta 1----2-D-xylose and alpha 1----3-L-fucose substituted N-linked oligosaccharides from Erythrina cristagalli lectin. Isolation, characterisation and comparison with other legume lectins. Eur J Biochem. 1987 Jul 15;166(2):311–320. doi: 10.1111/j.1432-1033.1987.tb13516.x. [DOI] [PubMed] [Google Scholar]
  2. Fournet B., Leroy Y., Wieruszeski J. M., Montreuil J., Poretz R. D., Goldberg R. Primary structure of an N-glycosidic carbohydrate unit derived from Sophora japonica lectin. Eur J Biochem. 1987 Jul 15;166(2):321–324. doi: 10.1111/j.1432-1033.1987.tb13517.x. [DOI] [PubMed] [Google Scholar]
  3. Hasilik A., Neufeld E. F. Biosynthesis of lysosomal enzymes in fibroblasts. Synthesis as precursors of higher molecular weight. J Biol Chem. 1980 May 25;255(10):4937–4945. [PubMed] [Google Scholar]
  4. Kaladas P. M., Goldberg R., Poretz R. D. Rabbit anti-carbohydrate antibody elicited by the lymphocyte mitogenic glycoprotein from Wistaria floribunda seeds. Mol Immunol. 1983 Jul;20(7):727–735. doi: 10.1016/0161-5890(83)90050-0. [DOI] [PubMed] [Google Scholar]
  5. Katz F., Moats W., Jan Y. N. A carbohydrate epitope expressed uniquely on the cell surface of Drosophila neurons is altered in the mutant nac (neurally altered carbohydrate). EMBO J. 1988 Nov;7(11):3471–3477. doi: 10.1002/j.1460-2075.1988.tb03222.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Kurosaka A., Yano A., Itoh N., Kuroda Y., Nakagawa T., Kawasaki T. The structure of a neural specific carbohydrate epitope of horseradish peroxidase recognized by anti-horseradish peroxidase antiserum. J Biol Chem. 1991 Mar 5;266(7):4168–4172. [PubMed] [Google Scholar]
  7. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  8. Laurière M., Laurière C., Chrispeels M. J., Johnson K. D., Sturm A. Characterization of a xylose-specific antiserum that reacts with the complex asparagine-linked glycans of extracellular and vacuolar glycoproteins. Plant Physiol. 1989 Jul;90(3):1182–1188. doi: 10.1104/pp.90.3.1182. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Mach L., Scherf W., Ammann M., Poetsch J., Bertsch W., März L., Glössl J. Purification and partial characterization of a novel lectin from elder (Sambucus nigra L.) fruit. Biochem J. 1991 Sep 15;278(Pt 3):667–671. doi: 10.1042/bj2780667. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Peumans W. J., Kellens J. T., Allen A. K., Van Damme E. J. Isolation and characterization of a seed lectin from elderberry (Sambucus nigra L.) and its relationship to the bark lectins. Carbohydr Res. 1991 Jun 25;213:7–17. doi: 10.1016/s0008-6215(00)90593-7. [DOI] [PubMed] [Google Scholar]
  11. Ramirez-Soto D., Poretz R. D. The (1----3)-linked alpha-L-fucosyl group of the N-glycans of the Wistaria floribunda lectins is recognized by a rabbit anti-serum. Carbohydr Res. 1991 Jun 25;213:27–36. doi: 10.1016/s0008-6215(00)90595-0. [DOI] [PubMed] [Google Scholar]
  12. Smith D. E., Fisher P. A. Identification, developmental regulation, and response to heat shock of two antigenically related forms of a major nuclear envelope protein in Drosophila embryos: application of an improved method for affinity purification of antibodies using polypeptides immobilized on nitrocellulose blots. J Cell Biol. 1984 Jul;99(1 Pt 1):20–28. doi: 10.1083/jcb.99.1.20. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Theisen N., Lohoff E. S., von Figura K., Hasilik A. Sequential detection of antigens in Western blots with differently colored products. Anal Biochem. 1986 Feb 1;152(2):211–214. doi: 10.1016/0003-2697(86)90399-4. [DOI] [PubMed] [Google Scholar]
  14. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Tretter V., Altmann F., März L. Peptide-N4-(N-acetyl-beta-glucosaminyl)asparagine amidase F cannot release glycans with fucose attached alpha 1----3 to the asparagine-linked N-acetylglucosamine residue. Eur J Biochem. 1991 Aug 1;199(3):647–652. doi: 10.1111/j.1432-1033.1991.tb16166.x. [DOI] [PubMed] [Google Scholar]
  16. Weber A., März L., Altmann F. Characteristics of the asparagine-linked oligosaccharide from honey-bee venom phospholipase A2. Evidence for the presence of terminal N-acetylglucosamine and fucose in an insect glycoprotein. Comp Biochem Physiol B. 1986;83(2):321–324. doi: 10.1016/0305-0491(86)90374-3. [DOI] [PubMed] [Google Scholar]
  17. Weber A., Schröder H., Thalberg K., März L. Specific interaction of IgE antibodies with a carbohydrate epitope of honey bee venom phospholipase A2. Allergy. 1987 Aug;42(6):464–470. doi: 10.1111/j.1398-9995.1987.tb00364.x. [DOI] [PubMed] [Google Scholar]
  18. Yasuda Y., Takahashi N., Murachi T. The composition and structure of carbohydrate moiety of stem bromelain. Biochemistry. 1970 Jan 6;9(1):25–32. doi: 10.1021/bi00803a004. [DOI] [PubMed] [Google Scholar]

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