Linkage of sugar chains to a fragment peptide of FGF-5S by a chemoenzymatic strategy and changes in the rate of proteolytic hydrolysis

被引:6
作者
Ajisaka, K
Miyasato, M
Ito, C
Fujita, Y
Yamazaki, Y
Oka, S
机构
[1] Meiji Milk Prod Co Ltd, Nutr Sci Inst, Kanagawa 2500862, Japan
[2] Natl Inst Adv Ind Sci & Technol, Tsukuba, Ibaraki 3058566, Japan
关键词
glycopeptide; enzymatic synthesis; FGF-5; proteolytic hydrolysis;
D O I
10.1023/A:1013613014830
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
Various O-linked and N-linked sugar chains were linked enzymatically to a fragment peptide (Leu-Ser-Gln(or Asn)-Val-His-Arg) of FGF-5S. First, galactose was linked with beta-(1-->3)-linkage to GalNAc-linked peptide by a transglycosylation using beta-galactosidase from Bacillus circulans (recombinant). Then sialic acid was linked with the aid of sialyltransferase from rat liver (recombinant) to give NeuAcalpha-(2-->3)-Galbeta-(1-->3)-GalNAc-linked hexapeptide. Further, a sialylated 2-chain biantennary sugar chain was linked by a transglycosylation using endo N-acetyl-beta-D-glucosaminidase from Mucor hiemalis (endo M, recombinant). The activity of DNA synthesis in a fibroblast cell line was increased by this glycosylation. The resistance of the obtained glycopeptides towards proteolytic hydrolysis by rat serum and by five proteases was compared with that of original peptide. The resistance was remarkably enhanced by the glycosylation.
引用
收藏
页码:301 / 308
页数:8
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