Synthesis and utilization of neo-glycoconjugates containing deaminoneuraminic acid (KDN) residues
含脱氨基神经氨酸(KDN)残基的新糖复合物的合成与利用
基本信息
- 批准号:07558211
- 负责人:
- 金额:$ 0.51万
- 依托单位:
- 依托单位国家:日本
- 项目类别:Grant-in-Aid for Scientific Research (B)
- 财政年份:1995
- 资助国家:日本
- 起止时间:1995 至 1996
- 项目状态:已结题
- 来源:
- 关键词:
项目摘要
The objective of this research project was to establish methods for enzymatic synthesis of neo-glycoconjugates containing new sialic acid (deaminoneuraminic acid, KDN) residues with the aim of utilization of such neo-KDN-glycoconjugates as new bioactive and/or organic materials. The following results were obtained :1. Search for KDN-specific glycosyltransferases and glycosidases-(1) Two KDN-transferases from rainbow trout ovary and sperm were found to be useful for neo-KDN-glycoconjugate synthesis ; -(2) Glycosidase highly specific for KDN residue was found and purified from a soil bacterium, and used for first identification of KDN-glycoconjugates in mammalian cells and tissues ;2. Establishment of methods for enzymatic synthesis of neo-KDN-glycoconjugates was achieved. Three synthetic processes using bacterial and animal enzymes were involved : synthesis of monosaccharide KDN from mannose and pyruvate with N-acylneuraminate lyase, synthesis of CMP-KDN from CTP and KDN with CMP-KDN synthetase, and incorporation of KDN residues into various glycoconjugates from CMP-KDN catalyzed by KDN-transferases. Using this method, glycoproteins, glycolipids, and oligosaccharides were able to be prepared in sufficient amounts for testing the biological and chemical properties. The enzymes involved in the latter two processes were discovered and prepared from rainbow trout by us.3. Neo-KDN-transferrin, prepared according to the methods described in 2, was examined for ligand activity to a plant lectin (SSA), which is known to be specific to NeuAc residue, revealing that SSA recognized KDN residue as well.4. For the purpose of in vivo synthesis of KDN-glycoconjugates, key enzyme(s) which determines KDN-glycoconjugate synthesis was searched for, and involvement of KDN 9-phosphate synthetase was suggested.
本研究项目的目的是建立酶促合成含有新的唾液酸(脱氨基神经氨酸,KDN)残基的新糖缀合物的方法,目的是利用这种新-KDN-糖缀合物作为新的生物活性和/或有机材料。获得了以下结果:1.寻找KDN-特异性糖基转移酶和糖苷酶-(1)发现来自虹鳟鱼卵巢和精子的两种KDN-转移酶可用于neo-KDN-糖缀合物的合成; -(2)从土壤细菌中发现并纯化了对KDN残基高度特异性的糖苷酶,并用于首次鉴定哺乳动物细胞和组织中的KDN-糖缀合物;2.建立了酶法合成neo-KDN-糖缀合物的方法。利用细菌和动物酶合成KDN涉及三个过程:用N-酰基神经氨酸裂解酶从甘露糖和丙酮酸合成单糖KDN,用CMP-KDN合成酶从CTP和KDN合成CMP-KDN,以及KDN转移酶催化CMP-KDN残基掺入各种糖缀合物中。使用该方法,能够制备足够量的糖蛋白、糖脂和寡糖以用于测试生物和化学性质。后两个过程所涉及的酶是我们从虹鳟鱼中发现并制备的。检查根据2中所述的方法制备的Neo-KDN-转铁蛋白对已知对NeuAc残基特异的植物凝集素(SSA)的配体活性,揭示SSA也识别KDN残基。为了在体内合成KDN-糖缀合物,寻找了决定KDN-糖缀合物合成的关键酶,并提出了KDN 9-磷酸合成酶的参与。
项目成果
期刊论文数量(18)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
Inoue, Sadako: "Identification of KDN residues in mammalalian tissues and human lung carcinoma cells." J. Biol. Cem.271. 24341-24344 (1996)
Inoue, Sadako:“哺乳动物组织和人肺癌细胞中 KDN 残基的鉴定。”
- DOI:
- 发表时间:
- 期刊:
- 影响因子:0
- 作者:
- 通讯作者:
Takaho Terada: "A new sialidase (KDNase Sm) catalysis initially forms a less stable anomer of KDN and is strongly inhibited by a transition-state analogue, KDN2en, but not by Neu2en5Ac" J.Biol.Chem.272(in press). (1997)
Takaho Terada:“一种新的唾液酸酶 (KDNase Sm) 催化作用最初形成一种不太稳定的 KDN 异头物,并被过渡态类似物 KDN2en 强烈抑制,但不会被 Neu2en5Ac 强烈抑制”J.Biol.Chem.272(出版中)。
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- 影响因子:0
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Takaho Terada: "Substrate specificity of rainbow trout testis CMP-KDN synthetase." Eur.J.Biochem.236. 852-855 (1996)
Takaho Terada:“虹鳟鱼睾丸 CMP-KDN 合成酶的底物特异性。”
- DOI:
- 发表时间:
- 期刊:
- 影响因子:0
- 作者:
- 通讯作者:
Inoue,Sadako: "Identificatton of KDN residues in mammalian tissues and human carcinoma cells." J.Biol.Chem.271. 24321-24344 (1996)
Inoue,Sadako:“哺乳动物组织和人类癌细胞中 KDN 残基的鉴定。”
- DOI:
- 发表时间:
- 期刊:
- 影响因子:0
- 作者:
- 通讯作者:
Terada, Takaho: "A new sialidase (KDNase Sm) catalysis initially forms a less stable anomer of KDN and is strongly inhibited by a transition-state analogue, KDN2en, but not by Neu2en5Ac." J. Biol. Chem.272 (in press). (1997)
Terada, Takaho:“一种新的唾液酸酶 (KDNase Sm) 催化作用最初形成一种不太稳定的 KDN 异头物,并受到过渡态类似物 KDN2en 的强烈抑制,但不受 Neu2en5Ac 的强烈抑制。”
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- 影响因子:0
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