Type analysis of the oligosaccharide chains on microheterogeneous components of bovine pancreatic DNAase by the lectin-nitrocellulose sheet method.
The oligosaccharide chains of microheterogeneous bovine pancreatic DNAases were characterized by the lectin-nitrocellulose sheet method. The active fractions of the DNAases from column chromatography showed four major and several minor spots on a two-dimensional polyacrylamide gel. They were transferred on to nitrocellulose sheets and treated with glycosidases (neuraminidase, endo-beta-N-acetyl glucosaminidase H or F, or peptide N-glycosidase F) and treated with peroxidase-coupled lectins (concanavalin A, Ricinus communis agglutinin or wheat-germ agglutinin). From the results, the most probable oligosaccharide types were proposed to be as follows: the four major spots contained components which had high-mannose type or hybrid-type oligosaccharides, such as those susceptible to endo-beta-N-acetylglucosaminidase H. In addition, spot 1 contained a complex-type biantennary oligosaccharide without sialic acid and spot 3 contained a tri- or tetra-antennary complex-type oligosaccharide with sialic acid. The component corresponding to spot 2 had a hybrid-type oligosaccharide chain with a 'bisecting' acetylglucosamine, linked 1-4 to the beta-mannose residue of the trimannosyl core, and the component corresponding to spot 4 had a high-mannose-type oligosaccharide chain.
Kijimoto-Ochiai
S
S
Institute of Immunological Science, Hokkaido University, Sapporo, Japan.
Katagiri
Y U
YU
Hatae
T
T
Okuyama
H
H
D000818:Animals; D002417:Cattle; D003101:Collodion; D003851:Deoxyribonucleases / Q000302:isolation & purification*; D015180:Electrophoresis, Gel, Two-Dimensional; D006026:Glycoside Hydrolases; D037102:Lectins; D008722:Methods; D009844:Oligosaccharides / Q000302:isolation & purification*; D010179:Pancreas / Q000201:enzymology*; D012794:Sialic Acids / Q000302:isolation & purification
D037102:Lectins; D009844:Oligosaccharides; D012794:Sialic Acids; D003101:Collodion; D003851:Deoxyribonucleases; D006026:Glycoside Hydrolases