The hexapeptide inhibitor of Galβ1,3GalNAc-specific α2,3-sialyltransferase as a generic inhibitor of sialyltransferases

被引:23
作者
Lee, KY
Kim, HG
Hwang, MR
Il Chae, J
Yang, JM
Lee, YC
Choo, YK
Lee, YI
Lee, SS
Do, SI
机构
[1] Korea Res Inst Biosci & Biotechnol, Cell Biol Div, Anim Cell & Med Glycobiol Lab, Taejon 305333, South Korea
[2] Korea Res Inst Biosci & Biotechnol, Liver Cell Signal Transduct Lab, Taejon 305333, South Korea
[3] Sogang Univ, Dept Life Sci, Seoul 100601, South Korea
[4] Dong A Univ, Div Nat Resources & Life Sci, Pusan 604022, South Korea
[5] WonKwang Univ, Div Biol Sci, Iksan 570749, South Korea
[6] Pai Chai Univ, Dept Biochem, Taejon 302735, South Korea
关键词
D O I
10.1074/jbc.M209618200
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 mammalian Galbeta1,3Ga1NAc-specific alpha2,3-sialyl-transferase (ST3Gal I) was expressed as a secreted glycoprotein in High Five(TM) (Trichoplusia ni) cells. Using this recombinant ST3Gal I, we screened the synthetic hexapeptide combinatorial library to explore a sialyltransferase inhibitor. We found that the hexapeptide, NH2-GNWWWW, exhibited the most strong inhibition of ST3Gal I among five different hexapeptides that were finally selected. The kinetic analysis of ST3Gal I inhibition demonstrated that this hexapeptide could act as a competitive inhibitor (K-i = 1.1 mum) on CMP-NeuAc binding to the enzyme. Moreover, the hexapeptide was shown to strongly inhibit both N-glycan-specific alpha2,3-and alpha2,6-sialyltranferase in vitro, suggesting that this peptide may inhibit the broad range of sialyltrans-ferases regardless of their linkage specificity. The inhibitory activity in vivo was investigated by RCA-I lectin blot analyses and by metabolic D-[6-H-3]GlcNH(2) radio-labeling analyses of N- and O-linked oligosaccharides in Chillies hamster ovary cells. Our results demonstrate that the hexapeptide can act as a generic inhibitor of the N- and O-glycan-specific sialyltransferases in mammalian cells, which results in the significantly reduced NeuAc expression on cellular glycoproteins in vivo.
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页码:49341 / 49351
页数:11
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