Differential expression of Galα1,3Gal epitope in polymeric and monomeric IgM secreted by mouse myeloma cells deficient in α2,6-sialyltransferase

Daniela Smilovich, Nadia Malagolini, Claudio Fagioli, Claudia De Lalla, Roberte Sitia, Franca Serafini-Cessi

Research output: Contribution to journalArticle

Abstract

IgM are glycoproteins secreted by plasma cells as (μ2L2)5+J or (μ2L2)6 polymers. In most species, μ- and J-chains bear five and one N-glycans, respectively. Here we compare the terminal glycosylation patterns of 4-hydroxy-3-nitrophenylacetyl (NP)-specific IgM secreted by transfectants of the J558L mouse myeloma deficient in the α2,6 sialyltransferase [α2,6ST(N)] or by a hybridoma expressing this enzyme (B1.8 cells). The absence of α2,6-sialylation results in an increased addition of α1,3-galactosyl residues to μ- and J-chain N-glycans. Since α1,3-galactosyltransferase (α1,3Gal-T) is similarly expressed in the two cell lines, these results indicate that a competition reaction occurs in vivo between α2,6ST(N) and α1,3Gal-T. In the α2,6ST(N) deficient transfectants, μ-chains lacking the C-subterminal Cys575 residue, which are secreted mainly in the form of μ2L2 monomers, are more efficiently capped by α1,3-galactosyl residues, confirming that polymerization significantly reduces the accessibility of μ-chain glycans to the Golgi processing enzymes involved in the biogenesis of antennary sugars. Functional assays indicate that IgM sialylation affects antigen-binding and complement-dependent hemolysis of haptenated red blood cells.

Original languageEnglish
Pages (from-to)841-848
Number of pages8
JournalGlycobiology
Volume8
Issue number8
DOIs
Publication statusPublished - Aug 1998

Fingerprint

Sialyltransferases
Polysaccharides
Immunoglobulin M
Epitopes
Cells
Glycosylation
Galactosyltransferases
Hybridomas
Enzymes
Hemolysis
Plasma Cells
Sugars
Polymerization
Assays
Glycoproteins
Polymers
Blood
Monomers
Erythrocytes
Plasmas

Keywords

  • α1,3 galactosyltransferase
  • α2,6 sialyltransferase-deficiency
  • IgM glycosylation
  • IgM polymerization

ASJC Scopus subject areas

  • Biochemistry

Cite this

Differential expression of Galα1,3Gal epitope in polymeric and monomeric IgM secreted by mouse myeloma cells deficient in α2,6-sialyltransferase. / Smilovich, Daniela; Malagolini, Nadia; Fagioli, Claudio; De Lalla, Claudia; Sitia, Roberte; Serafini-Cessi, Franca.

In: Glycobiology, Vol. 8, No. 8, 08.1998, p. 841-848.

Research output: Contribution to journalArticle

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N2 - IgM are glycoproteins secreted by plasma cells as (μ2L2)5+J or (μ2L2)6 polymers. In most species, μ- and J-chains bear five and one N-glycans, respectively. Here we compare the terminal glycosylation patterns of 4-hydroxy-3-nitrophenylacetyl (NP)-specific IgM secreted by transfectants of the J558L mouse myeloma deficient in the α2,6 sialyltransferase [α2,6ST(N)] or by a hybridoma expressing this enzyme (B1.8 cells). The absence of α2,6-sialylation results in an increased addition of α1,3-galactosyl residues to μ- and J-chain N-glycans. Since α1,3-galactosyltransferase (α1,3Gal-T) is similarly expressed in the two cell lines, these results indicate that a competition reaction occurs in vivo between α2,6ST(N) and α1,3Gal-T. In the α2,6ST(N) deficient transfectants, μ-chains lacking the C-subterminal Cys575 residue, which are secreted mainly in the form of μ2L2 monomers, are more efficiently capped by α1,3-galactosyl residues, confirming that polymerization significantly reduces the accessibility of μ-chain glycans to the Golgi processing enzymes involved in the biogenesis of antennary sugars. Functional assays indicate that IgM sialylation affects antigen-binding and complement-dependent hemolysis of haptenated red blood cells.

AB - IgM are glycoproteins secreted by plasma cells as (μ2L2)5+J or (μ2L2)6 polymers. In most species, μ- and J-chains bear five and one N-glycans, respectively. Here we compare the terminal glycosylation patterns of 4-hydroxy-3-nitrophenylacetyl (NP)-specific IgM secreted by transfectants of the J558L mouse myeloma deficient in the α2,6 sialyltransferase [α2,6ST(N)] or by a hybridoma expressing this enzyme (B1.8 cells). The absence of α2,6-sialylation results in an increased addition of α1,3-galactosyl residues to μ- and J-chain N-glycans. Since α1,3-galactosyltransferase (α1,3Gal-T) is similarly expressed in the two cell lines, these results indicate that a competition reaction occurs in vivo between α2,6ST(N) and α1,3Gal-T. In the α2,6ST(N) deficient transfectants, μ-chains lacking the C-subterminal Cys575 residue, which are secreted mainly in the form of μ2L2 monomers, are more efficiently capped by α1,3-galactosyl residues, confirming that polymerization significantly reduces the accessibility of μ-chain glycans to the Golgi processing enzymes involved in the biogenesis of antennary sugars. Functional assays indicate that IgM sialylation affects antigen-binding and complement-dependent hemolysis of haptenated red blood cells.

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