IDENTIFICATION OF PEPTIDE-N-GLYCANASE ACTIVITY IN MAMMALIAN DERIVED CULTURED-CELLS

被引:92
作者
SUZUKI, T [1 ]
SEKO, A [1 ]
KITAJIMA, K [1 ]
INOUE, Y [1 ]
INOUE, S [1 ]
机构
[1] SHOWA UNIV,SCH PHARMACEUT SCI,TOKYO 142,JAPAN
关键词
D O I
10.1006/bbrc.1993.1938
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 recent finding of peptide:N-glycanase (PNGase) in medaka embryos (Seko, A., Kitajima, K., Inoue, Y., & Inoue, S., J.Biol. Chem. 266, 22110 (1991)) raised the question of how widespread is the occurrence of this type of de-N-glycosylating enzyme. In experiments designed to identify PNGase in the mammalian system, we searched for its activity in some cultured cell lines. Incubation of a 14C-labeled N-glycopeptide with extracts prepared from cultured cells resulted in producing the free glycan and the peptide. Detailed characterizations of the products, formed upon incubation of a 14C-labeled N-glycopeptide substrate with the enzyme preparation from C3H mouse loose connective tissue-derived L-929 cells, by HPLC, amino acid and carbohydrate composition analyses, and peptide sequence analysis unequivocally established the reaction products to be the free glycan having di-N-acetylchitobiosyl sequence at its reducing end and free peptide in which the originally glycan-linked Asn residue was converted to the Asp residue. This represents the first demonstration of PNGase in mammalian cells and thus PNGase appears to be a very common enzyme expressed in not only plants and bacteria but also a wide range of animals although its functional significance remains to be clarified. © 1993 Academic Press. All rights reserved.
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页码:1124 / 1130
页数:7
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