Development ofenzyme inlulkar far infectious diseace including malaria and clarification ofthe inlubitor activity

疟疾等远距离传染病酶的研制及其抑制剂活性的阐明

• 批准号: 18350086
• 负责人: INOUE Tsuyoshi
• 金额: $ 7.92万
• 依托单位: Osaka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2006
• 资助国家: 日本
• 起止时间: 2006 至 2007
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18350086/
• 关键词: Plasmodium falciparum; infectious desease; drug design; X-ray structure analysis

Tropical diseases such as human malaria, leishmaniasis, Chagas disease, human African trypanosomiasis, dengue fever and schistosomiasis continue to cause significant morbidity and mortality. In this project, we performed X-ray crystallographic studies of important enzymes from parasite concerning three these infectious disease to construct the structural basis for the drug development by the in-silica method. The important enzymes are (1) two kinds of enzymes concerning the de novo synthesis of pyrimidine nucleotide from human malaria, (2) prostaglandin (PG) F synthases from parasites of Chagas' disease and Leishmania.Recombinant enzymes of orotate phosphoribosyltransferase (pyr5) and Orotidine 5'-monophosphate decarboxylase (pyr6) were purified from E.coli and crystallized. As the result, the native and product UMP complex form of pyr6 were crystallized and carried out with the X-ray structural analyses. The 2.7 and 2.6 A resolutional structures of apo- and complex forms of pyr6 provided the details of the decarboxylation mechanism (J. Biochem., 2008). As for the PGF synthases from Trypanosoma cruzi and Leishmania mejor were also performed crystallization and the structure of PGF synthase from T. cruzi were determined at 1.7 A resolution (Acta Cryst. F, 2007). The complex structure of PGF sytnthase with indomethacin were also determined, providing the structural information for new inhibitor development.

人类疟疾、利什曼病、恰加斯病、非洲锥虫病、登革热和血吸虫病等热带疾病继续造成严重的发病率和死亡率。在这个项目中，我们对与这三种传染病有关的寄生虫中的重要酶进行了 X 射线晶体学研究，为通过硅内方法开发药物奠定了结构基础。重要的酶是（1）与人类疟疾中的嘧啶核苷酸从头合成有关的两种酶，（2）来自南美锥虫病和利什曼原虫寄生虫的前列腺素（PG）F合酶。乳清酸磷酸核糖基转移酶（pyr5）和乳清酸核苷5'-单磷酸的重组酶 从大肠杆菌中纯化脱羧酶 (pyr6) 并结晶。结果，pyr6 的天然和产物 UMP 复合物形式被结晶并进行了 X 射线结构分析。 Pyr6 的脱辅基形式和复合形式的 2.7 和 2.6 A 解析结构提供了脱羧机制的详细信息 (J. Biochem., 2008)。对于来自克氏锥虫和大利什曼原虫的 PGF 合酶也进行了结晶，并且以 1.7 A 分辨率测定了来自克氏锥虫的 PGF 合酶的结构（Acta Cryst.F，2007）。还确定了PGF合酶与吲哚美辛的复合结构，为新抑制剂的开发提供了结构信息。

项目成果

Structural basis for the decarboxylation of orotidine 5'-monophosphate (OMP) by Plasmodium Talon : Far= OMP decarboxylase.

疟原虫 Talon 使乳清苷 5-单磷酸 (OMP) 脱羧的结构基础：Far= OMP 脱羧酶。

• 发表时间: 2008
• 期刊: The Journal of Biochemistry 143
• 作者: K.;Tokuoka;et. al.
• 通讯作者: et. al.

Trypanosoma cruzi由来01d Yellow EnzymeのX線結晶構造解析

克氏锥虫 01d 黄色酶的 X 射线晶体结构分析

• 发表时间: 2007
• 作者: N.;Okamoto;H. Okano;岡本尚毅
• 通讯作者: 岡本尚毅

The Crystal Structure of Orotidine 5'-monophosphate Decarboxylase from the Human Malaria Parasite Plasmodium falciparm

人疟原虫恶性疟原虫乳清苷 5-单磷酸脱羧酶的晶体结构

• 发表时间: 2008
• 作者: K.;Tokuoka
• 通讯作者: Tokuoka

Crystal Structure analysis of Prostaglandin F_<2a> synthase.

前列腺素F_<2a>合酶的晶体结构分析。

• 发表时间: 2007
• 作者: N.;Okamoto;H. Okano;岡本尚毅;徳岡啓司
• 通讯作者: 徳岡啓司

Crystallization and preliminary X-ray analysis of the tRNA thiolation enzyme MnmA from Escherichia coli complexed with tRNA(Glu)

大肠杆菌 tRNA 硫醇化酶 MnMA 与 tRNA(Glu) 复合物的结晶和初步 X 射线分析

• 发表时间: 2006
• 期刊: Acta Crystallograph Sect F Struct Biol Cryst Commun 62
• 作者: Numata;T.;Ikeuchi;Y.;Fukai;S.;Adachi;H.;Matsumura;H.;Takano;K.;Murakami;S.;Inoue;T.;Mori;Y.;Sasaki;T.;Suzuki;T.;Nureki;O.
• 通讯作者: O.