Biosynthesis of mammary glycoproteins. Partial characterization of the sequence for the assembly of lipid-linked saccharides

I. K. Vijay, Gary H. Perdew, D. E. Lewis

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Abstract

Incubation of a membrane preparation from the lactating bovine mammary gland with UDP-[3H]GlcNAc, GDP-[14C]Man, and UDP-[3H]Glc results in the biosynthesis of 15 lipid-linked saccharides that differ from one another by a monosaccharide unit. Pulse and chase kinetics indicate that these glycolipids are related to one another as precursor products for the biosynthesis of asparagine-linked glycoproteins of this tissue. [Man-14C]- and [Man-14C, GlcNAc-3H]saccharides were prepared from corresponding glycolipids by mild acid hydrolysis. . Following extensive purification by paper and gel filtration chromatography, structural characterization was conducted on tri-, tetra-, penta-, and undecasaccharides via size determination on calibrated columns of Bio-Gel P-2 and P-4, compositional analysis, exo- and endoglycosidase digestions, methylation, Smith degradation, and acetolysis. These structures were identified as: Manβ1→4(3)GlcNacβ1→4(3)Glc-Nac, Manα1→3Manβ1→4(3)GlcNAcβ1→4(3)GlcNAc, Manα1→3(Manα1→6)Manβ1→4(3)GlcNAcβ1→4(3)Glc-NAc, and Manα1→2Manα1→2Manα1→3(Manα1→2Manα1→6[Manα1→2Manα1→3]Manα1→6)Manβ1→4(3)GlcNAcβ1→4(3)GlcNAc.

Original languageEnglish (US)
Pages (from-to)11210-11220
Number of pages11
JournalJournal of Biological Chemistry
Volume255
Issue number23
StatePublished - Dec 1 1980

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All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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