Carbohydrates of lysosomal enzymes secreted by Tetrahymena pyriformis.
The carbohydrate structures of acid phosphatase and alpha-glucosidase secreted into culture medium by Tetrahymena pyriformis strain W were studied. Their asparagine-linked sugar chains were quantitatively liberated as radioactive oligosaccharides from their polypeptide moieties by controlled hydrazinolysis followed by N-acetylation and NaB3H4 reduction. The approximate amounts of total sugar chains liberated from 1 mol each of acid phosphatase and alpha-glucosidase were 6 and 4 mol, respectively. Paper electrophoresis revealed that only neutral oligosaccharides were obtained from both enzymes. The oligosaccharide fraction from acid phosphatase was separated into seven components by Bio-Gel P-4 column chromatography while that from alpha-glucosidase was resolved into three components. The structures of these oligosaccharides were determined by sequential glycosidase digestion in combination with methylation analysis. The sugar chains of the two enzymes can be primarily classified as high mannose-type oligosaccharides. However, they have the following characteristic features: 1) their common core is not the usual Man5 . GlcNAc2 structure, it is Man3 . GlcNAc2; 2) some of the sugar chains of acid phosphatase have 1 approximately 3 glucose residues linked to the nonreducing terminal Man alpha 1----2 residue. The structural characteristics of the sugar moieties of the two enzymes indicate that they might be produced by the so-called "alternate pathway," in which lipid-linked Glc3 . Man5 . GlcNAc2 functions as an oligosaccharide donor.
Taniguchi
T
T
Mizuochi
T
T
Banno
Y
Y
Nozawa
Y
Y
Kobata
A
A
D000135:Acid Phosphatase / Q000032:analysis*; D000818:Animals; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D005959:Glucosidases / Q000032:analysis*; D006026:Glycoside Hydrolases / Q000378:metabolism; D006596:Hexosaminidases / Q000378:metabolism; D008247:Lysosomes / Q000201:enzymology*; D008361:Mannosidases / Q000378:metabolism; D008745:Methylation; D009844:Oligosaccharides / Q000032:analysis* / Q000378:metabolism; D013769:Tetrahymena pyriformis / Q000201:enzymology*; D000520:alpha-Glucosidases / Q000032:analysis* / Q000378:metabolism; D001616:beta-Galactosidase / Q000378:metabolism; D001619:beta-N-Acetylhexosaminidases
D009844:Oligosaccharides; D000135:Acid Phosphatase; D005959:Glucosidases; D006026:Glycoside Hydrolases; D006596:Hexosaminidases; D008361:Mannosidases; D000520:alpha-Glucosidases; D001616:beta-Galactosidase; D001619:beta-N-Acetylhexosaminidases