Identification of methylphosphomannosyl residues as components of the high mannose oligosaccharides of Dictyostelium discoideum glycoproteins.
Lysosomal enzymes isolated from the slime mold Dictyostelium discoideum bind to the mannose 6-phosphate receptor which is present in many mammalian cells. While binding to the receptor suggests that the slime mold enzymes possess the same mannose 6-phosphate recognition marker as their mammalian counterparts, initial structural studies of the phosphorylated oligosaccharides have indicated that the phosphate is attached to high mannose-type units via an unusual phosphodiester linkage (Freeze, H.H., Yeh, R., Miller, A.L., and Kornfeld, S. (1983) J. Biol. Chem. 258, 14874-14879). To identify the components of the phosphodiester group we have isolated the phosphorylated high-mannose oligosaccharides from D. discoideum AX3 cells labeled with [2-3H]mannose or [6-3H]glucosamine and from the differentiation medium of unlabeled cells. The major phosphorylated species contain one or two phosphodiester groups and an average of 6 or 7 mannose residues. The phosphodiesters are relatively stable to both acid and base hydrolysis, but upon strong acid hydrolysis (conditions that completely hydrolyze the oligosaccharide) mannose 6-phosphate residues are liberated. Through a combination of techniques, including fast atom bombardment and direct chemical ionization mass spectrometry, it is shown that the mannose 6-phosphate residues of the intact oligosaccharide are diesterified to methyl groups. This indicates that slime mold possesses a different biosynthetic pathway for the formation of phosphorylated high mannose-type oligosaccharides than is utilized by higher organisms.
D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D002854:Chromatography, Paper; D004023:Dictyostelium / Q000032:analysis*; D006600:Hexosephosphates / Q000032:analysis*; D009682:Magnetic Resonance Spectroscopy; D008360:Mannosephosphates / Q000032:analysis*; D013058:Mass Spectrometry; D008759:Methylglycosides / Q000032:analysis*; D008766:Methylmannosides / Q000032:analysis*; D009844:Oligosaccharides / Q000032:analysis*
D006600:Hexosephosphates; D008360:Mannosephosphates; D008759:Methylglycosides; D008766:Methylmannosides; D009844:Oligosaccharides