Glycolipid precursors for the membrane anchor of Trypanosoma brucei variant surface glycoproteins.

I. Can structure of the phosphatidylinositol-specific phospholipase C sensitive and resistant glycolipids

Satyajit Mayor, Anant K. Menon, George A. M. Cross, Michael A. J. Ferguson, Raymond A. Dwek, Thomas W. Rademacher

    Research output: Contribution to journalArticle

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    Abstract

    A number of eukaryotic surface glycoproteins, including the variant surface glycoproteins of Trypanosoma brucei, are synthesized with a carboxyl-terminal hydrophobic peptide extension that is cleaved and replaced by a complex glycosyl-phosphatidylinositol (GPI) membrane anchor within 1-5 min of the completion of polypeptide synthesis. The rapidity of this carboxyl-terminal modification suggests the existence of a prefabricated precursor glycolipid that can be transferred en bloc to the polypeptide. We have reported the purification and partial characterization of a candidate precursor glycolipid (P2) and of a compositionally similar glycolipid (P3) from T. brucei (Menon, A.K., Mayor, S., Ferguson, M.A.J., Duszenko, M., and Cross, G.A.M. (1988) J. Biol. Chem. 263, 1970-1977). The primary structure of the glycan portions of P2 and P3 have now been analyzed by a combination of selective chemical fragmentation and enzymatic glycan sequencing at the subnanomolar level. The glycans were generated by deamination, NaB3H4 reduction, and dephosphorylation of glycolipids purified from different trypanosome variants. Glycan fragments derived from biosynthetically labeled glycolipids were also analyzed. The cumulative data strongly suggest that P2 and P3 contain ethanolamine-phosphate-Mana1-2Mana1-6Mana1-GlcN linked glycosidically to an inositol residue, as do all the GPI anchors that have been structurally characterized. The structural similarities suggest that GPI membrane anchors are derived from common precursor glycolipids that become variably modified during or after addition to newly synthesized proteins.

    Original languageEnglish
    Pages (from-to)6164-6173
    Number of pages10
    JournalJournal of Biological Chemistry
    Volume265
    Issue number11
    Publication statusPublished - 15 Apr 1990

    Fingerprint

    Trypanosoma Variant Surface Glycoproteins
    Phosphoinositide Phospholipase C
    Trypanosoma brucei brucei
    Glycolipids
    Membrane Glycoproteins
    Anchors
    Membranes
    Glycosylphosphatidylinositols
    Polysaccharides
    Peptides
    Deamination
    Trypanosomiasis
    Inositol
    Purification

    Cite this

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    title = "Glycolipid precursors for the membrane anchor of Trypanosoma brucei variant surface glycoproteins.: I. Can structure of the phosphatidylinositol-specific phospholipase C sensitive and resistant glycolipids",
    abstract = "A number of eukaryotic surface glycoproteins, including the variant surface glycoproteins of Trypanosoma brucei, are synthesized with a carboxyl-terminal hydrophobic peptide extension that is cleaved and replaced by a complex glycosyl-phosphatidylinositol (GPI) membrane anchor within 1-5 min of the completion of polypeptide synthesis. The rapidity of this carboxyl-terminal modification suggests the existence of a prefabricated precursor glycolipid that can be transferred en bloc to the polypeptide. We have reported the purification and partial characterization of a candidate precursor glycolipid (P2) and of a compositionally similar glycolipid (P3) from T. brucei (Menon, A.K., Mayor, S., Ferguson, M.A.J., Duszenko, M., and Cross, G.A.M. (1988) J. Biol. Chem. 263, 1970-1977). The primary structure of the glycan portions of P2 and P3 have now been analyzed by a combination of selective chemical fragmentation and enzymatic glycan sequencing at the subnanomolar level. The glycans were generated by deamination, NaB3H4 reduction, and dephosphorylation of glycolipids purified from different trypanosome variants. Glycan fragments derived from biosynthetically labeled glycolipids were also analyzed. The cumulative data strongly suggest that P2 and P3 contain ethanolamine-phosphate-Mana1-2Mana1-6Mana1-GlcN linked glycosidically to an inositol residue, as do all the GPI anchors that have been structurally characterized. The structural similarities suggest that GPI membrane anchors are derived from common precursor glycolipids that become variably modified during or after addition to newly synthesized proteins.",
    author = "Satyajit Mayor and Menon, {Anant K.} and Cross, {George A. M.} and Ferguson, {Michael A. J.} and Dwek, {Raymond A.} and Rademacher, {Thomas W.}",
    year = "1990",
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    pages = "6164--6173",
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    Glycolipid precursors for the membrane anchor of Trypanosoma brucei variant surface glycoproteins. I. Can structure of the phosphatidylinositol-specific phospholipase C sensitive and resistant glycolipids. / Mayor, Satyajit; Menon, Anant K.; Cross, George A. M.; Ferguson, Michael A. J.; Dwek, Raymond A.; Rademacher, Thomas W.

    In: Journal of Biological Chemistry, Vol. 265, No. 11, 15.04.1990, p. 6164-6173.

    Research output: Contribution to journalArticle

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    T2 - I. Can structure of the phosphatidylinositol-specific phospholipase C sensitive and resistant glycolipids

    AU - Mayor, Satyajit

    AU - Menon, Anant K.

    AU - Cross, George A. M.

    AU - Ferguson, Michael A. J.

    AU - Dwek, Raymond A.

    AU - Rademacher, Thomas W.

    PY - 1990/4/15

    Y1 - 1990/4/15

    N2 - A number of eukaryotic surface glycoproteins, including the variant surface glycoproteins of Trypanosoma brucei, are synthesized with a carboxyl-terminal hydrophobic peptide extension that is cleaved and replaced by a complex glycosyl-phosphatidylinositol (GPI) membrane anchor within 1-5 min of the completion of polypeptide synthesis. The rapidity of this carboxyl-terminal modification suggests the existence of a prefabricated precursor glycolipid that can be transferred en bloc to the polypeptide. We have reported the purification and partial characterization of a candidate precursor glycolipid (P2) and of a compositionally similar glycolipid (P3) from T. brucei (Menon, A.K., Mayor, S., Ferguson, M.A.J., Duszenko, M., and Cross, G.A.M. (1988) J. Biol. Chem. 263, 1970-1977). The primary structure of the glycan portions of P2 and P3 have now been analyzed by a combination of selective chemical fragmentation and enzymatic glycan sequencing at the subnanomolar level. The glycans were generated by deamination, NaB3H4 reduction, and dephosphorylation of glycolipids purified from different trypanosome variants. Glycan fragments derived from biosynthetically labeled glycolipids were also analyzed. The cumulative data strongly suggest that P2 and P3 contain ethanolamine-phosphate-Mana1-2Mana1-6Mana1-GlcN linked glycosidically to an inositol residue, as do all the GPI anchors that have been structurally characterized. The structural similarities suggest that GPI membrane anchors are derived from common precursor glycolipids that become variably modified during or after addition to newly synthesized proteins.

    AB - A number of eukaryotic surface glycoproteins, including the variant surface glycoproteins of Trypanosoma brucei, are synthesized with a carboxyl-terminal hydrophobic peptide extension that is cleaved and replaced by a complex glycosyl-phosphatidylinositol (GPI) membrane anchor within 1-5 min of the completion of polypeptide synthesis. The rapidity of this carboxyl-terminal modification suggests the existence of a prefabricated precursor glycolipid that can be transferred en bloc to the polypeptide. We have reported the purification and partial characterization of a candidate precursor glycolipid (P2) and of a compositionally similar glycolipid (P3) from T. brucei (Menon, A.K., Mayor, S., Ferguson, M.A.J., Duszenko, M., and Cross, G.A.M. (1988) J. Biol. Chem. 263, 1970-1977). The primary structure of the glycan portions of P2 and P3 have now been analyzed by a combination of selective chemical fragmentation and enzymatic glycan sequencing at the subnanomolar level. The glycans were generated by deamination, NaB3H4 reduction, and dephosphorylation of glycolipids purified from different trypanosome variants. Glycan fragments derived from biosynthetically labeled glycolipids were also analyzed. The cumulative data strongly suggest that P2 and P3 contain ethanolamine-phosphate-Mana1-2Mana1-6Mana1-GlcN linked glycosidically to an inositol residue, as do all the GPI anchors that have been structurally characterized. The structural similarities suggest that GPI membrane anchors are derived from common precursor glycolipids that become variably modified during or after addition to newly synthesized proteins.

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