Site-specific glycosylation of bovine butyrophilin.
Our previous studies showed that the N-linked sugar chains of most bovine glycoproteins from milk fat globule membranes (MFGM) contain the GalNAc beta 1-->4GlcNAc group [Sato et al. (1993) J. Biochem. 114, 890-900]. Since expression of the disaccharide structure is influenced by peptide sequences near the glycosylation sites [Smith and Baenziger (1992) Proc. Natl. Acad. Sci. USA 89, 329-333], the site-specificity of the N-acetylgalactosaminylated sugar chains was investigated using bovine butyrophilin, a major MFGM glycoprotein with known primary structure. Two glycopeptide fragments which contained the N-linked sugar chains linked to either Asn-55 or Asn-215 residue were obtained by digestion of the protein with Achromobacter protease I. The sugar chains released from each glycopeptide by hydrazinolysis were reduced with NaB3H4. Structural analyses of the oligosaccharides by sequential exoglycosidase digestion and methylation analysis revealed that only complex-type sugar chains with the GalNAc beta 1-->4GlcNAc structure are included in Asn-55-linked oligosaccharides, while only novel hybrid-type sugar chains detected previously in bovine MFGM glycoproteins are included in Asn-215-linked oligosaccharides. The results show that the glycosylation of butyrophilin occurs in a site-specific manner.
Sato
T
T
Department of Biochemistry, Institute of Medical Science, University of Tokyo.
Takio
K
K
Kobata
A
A
Greenwalt
D E
DE
Furukawa
K
K
D000595:Amino Acid Sequence; D000818:Animals; D000071797:Butyrophilins; D002240:Carbohydrate Sequence; D002417:Cattle; D002846:Chromatography, Affinity; D004586:Electrophoresis; D006031:Glycosylation; D008562:Membrane Glycoproteins / Q000737:chemistry*; D008745:Methylation; D008894:Milk Proteins / Q000737:chemistry*; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000302:isolation & purification*; D010446:Peptide Fragments / Q000302:isolation & purification*
D000071797:Butyrophilins; D008562:Membrane Glycoproteins; D008894:Milk Proteins; D009844:Oligosaccharides; D010446:Peptide Fragments