硫酸乙酰肝素的表达D-glucosaminyl 3-O-sulfotransferase亚型揭示小说底物特异性。
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刘J, Shworak NW, Sinay P,施瓦茨JJ,张L Fritze LM,罗森博格RD
硫酸乙酰肝素的表达D-glucosaminyl 3-O-sulfotransferase亚型揭示小说底物特异性。
生物化学杂志。1999年2月19日,274 (8):5185 - 92。
- PubMed ID
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9988768 (在PubMed]
- 文摘
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葡萄糖胺残留的3-O-sulfation硫酸乙酰肝素的生物合成中是一个重要的修改(HS)。我们先前的研究让我们净化和硫酸乙酰肝素分子克隆D-glucosaminyl 3-O-sulfotransferase (3-OST-1),即关键酶转换nonanticoagulant硫酸乙酰肝素(HSinact)抗凝剂硫酸乙酰肝素(HSact)。在这项研究中,我们表示和特征3-OST-1同源基因的全长cdna,指定为3-OST-2, 3-OST-3A, 3-OST-3B中描述相应的论文(Shworak: W。刘,J。佩,l . M。张,L。小林,M。科普兰,n G。詹金斯,n。罗森博格,r . d .(1999)生物。274年化学,5170 - 5184)。所有这些互补在COS-7细胞成功表达,和硫酸乙酰肝素sulfotransferase活动被发现在细胞中提取。我们证明3-OST-2 3-OST-3A, 3-OST-3B硫酸乙酰肝素D-glucosaminyl 3-O-sulfotransferases因为硫酸转移酶腺苷3”-phosphophate 5“磷-硫酸35 [s] ([35 s] PAPS) 3-OH葡萄糖胺的位置。 3-OST-3A and 3-OST-3B sulfate an identical disaccharide. HSact conversion activity in the cell extract transfected by 3-OST-1 was shown to be 300-fold greater than that in the cell extracts transfected by 3-OST-2 and 3-OST-3A, suggesting that 3-OST-2 and 3-OST-3A do not make HSact. The results of the disaccharide analysis of the nitrous acid-degraded [35S]HS suggested that 3-OST-2 transfers sulfate to GlcA2S-GlcNS and IdoA2S-GlcNS; 3-OST-3A transfers sulfate to IdoA2S-GlcNS. Our results demonstrate that the 3-O-sulfation of glucosamine is generated by different isoforms depending on the saccharide structures around the modified glucosamine residue. This discovery has provided evidence for a new cellular mechanism for generating a defined saccharide sequence in structurally complex HS polysaccharide.