Detection of Man5GlcNAc and related free oligomannosides in the cytosol fraction of hen oviduct

被引:30
作者
Iwai, K [1 ]
Mega, T [1 ]
Hase, S [1 ]
机构
[1] Osaka Univ, Grad Sch Sci, Dept Chem, Toyonaka, Osaka 5600043, Japan
关键词
cytosol; free oligosaccharides; hen oviduct; oligomannoside;
D O I
10.1093/oxfordjournals.jbchem.a022270
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The presence of free oligomannosides in cytosol has been demonstrated by metabolic radiolabeling, and Man alpha 1-2Man alpha 1-2Man alpha 1-3(Man alpha 1-6)Man beta 1-4GlcNAc was detected as the main free oligomannoside in Chinese hamster ovary cells [Kmiecik, D., Herman, V., Stroop, C.J.M., Michalski, J.C., Mir, A.M., Labiau, O., Verbert, A., and Cacan, R. (1995) Glycobiology, 5, 483-494]. In the present paper, the isomeric structures and amounts of oligomannosides in the cytosol fraction of hen oviduct were analyzed by pyridylamination and exoglycosidase digestion. Hen oviduct was used since our group has already characterized the cytosolic neutral alpha-mannosidase and endo-beta-N-acetylglucosaminidase obtained from the same source. The amounts of Man(2)GlcNAc, Man(3)GlcNAc, Man(4)GlcNAc, and Man(5)GlcNAc were 0.6, 0.6, 0.5, and 0.8nmol/g tissue, respectively, but Man(6)GlcNAc-Man(9) GlcNAc were not detected. The isomeric structures of the Man(3)GlcNAc-Man(5)GlcNAc found were (Man alpha 1-2)(0-2)Man alpha 1-3(Man alpha 1-6)Man beta 1-4GlcNAc, which were compatible with the substrate specificities of cytosolic endo-beta-N-acetylglucosaminidase and neutral alpha-mannosidase, indicating that these enzymes participate in the formation of the oligomannosides present in the cytosol.
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收藏
页码:70 / 74
页数:5
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