Product release mechanism of Lipocalin-type Prostaglandin D synthase

脂质运载蛋白型前列腺素D合酶的产物释放机制

• 批准号: 24790055
• 负责人: SHIMAMOTO Shigeru
• 金额: $ 2.83万
• 依托单位: Kinki University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Young Scientists (B)
• 财政年份: 2012
• 资助国家: 日本
• 起止时间: 2012-04-01 至 2014-03-31
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-24790055/
• 关键词: リポカリン型プロスタグランジンD合成酵素; プロスタグランジンD2; NMR; ITC; プロスタグランジンD

Lipocalin-type Prostaglandin D synthase (L-PGDS) catalyzes the isomerization of prostaglandin H2 (PGH2) to produce prostaglandin D2 (PGD2), which acts as a potent endogenous somnogen in the brain. A number of studies of L-PGDS, as a drug target for sleep disorders, have been reported in attempts to understand its catalytic mechanism, and several substrate recognition models of L-PGDS have been proposed. However, details of the mechanism by which L-PDGS recognizes its substrates and products are obscure, since essential information, such as its binding affinity and stoichiometry, of the interactions between L-PGDS and its substrates and products remains unclear. Therefore, we carried out ITC and NMR experiments to characterize the binding properties. The results of the ITC and NMR measurements revealed that both the substrate and the product bind to L-PGDS in a stoichiometry of 2 to 1 and two binding sites, namely a high and a low affinity binding site, are present.

Lipocalin型前列腺素D合成酶(L-PGDS)催化前列腺素H2(PGH2)异构化生成前列腺素D2(PGD2)，前列腺素D2是脑内一种强有力的内源性睡眠激素。L-前列腺素DS作为治疗睡眠障碍的药物靶点已有大量研究报道，试图了解其催化机制，并提出了几种L-PGDS的底物识别模型。然而，L-PDGS识别其底物和产物的详细机制尚不清楚，因为L-PGDS与其底物和产物相互作用的基本信息，如其结合亲和力和化学计量比仍不清楚。因此，我们进行了ITC和核磁共振实验来表征其结合性质。红外光谱和核磁共振结果表明，底物和产物均以2：1的化学计量比与L-PGDS结合，并存在两个结合部位，即高亲和力结合部位和低亲和力结合部位。

项目成果

(1)H, (13)C, and (15)N resonance assignments of mouse lipocalin-type prostaglandin D synthase/substrate analog complex.

小鼠脂质运载蛋白型前列腺素 D 合酶/底物类似物复合物的 (1)H、(13)C 和 (15)N 共振归属。

• DOI: 10.1007/s12104-013-9467-5
• 发表时间: 2014
• 期刊: Biomolecular NMR Assignments
• 影响因子: 0.9
• 作者: Shimamoto Shigeru;Maruo Hiroko;Yoshida Takuya;Ohkubo Tadayasu.
• 通讯作者: Ohkubo Tadayasu.

Chemical methods and approaches to the regioselective formation of multiple disulfide bonds

区域选择性形成多个二硫键的化学方法和途径

• 发表时间: 2014
• 期刊: Curr. Protoc. Protein Sci
• 作者: Shimamoto S.;Katayama H.;Okumura M.;and Hidaka Y
• 通讯作者: and Hidaka Y

Chemical Assistance of Disulfide Coupled Peptide and Protein Folding

二硫键偶联肽和蛋白质折叠的化学辅助

• 发表时间: 2014
• 期刊: Curr. Protoc. Protein Sci
• 作者: Okumura M.;Shimamoto S.;and Hidaka Y
• 通讯作者: and Hidaka Y

リポカリン型プロスタグランジンD合成酵素と基質・生成物の相互作用解析

脂质运载蛋白型前列腺素D合酶与底物/产物的相互作用分析

• 发表时间: 2014
• 作者: 福田裕太郎（発表者）;島本 茂;丸野孝浩;小林祐次;大久保忠恭;有竹浩介; 裏出良博;日髙雄二（責任者）
• 通讯作者: 日髙雄二（責任者）

STRUCTURAL AND DYNAMIC ANALYSIS OF LIPOCALIN TYPE PROSTAGLANDIN D SYNTHASE IN ITS APO FORM AND SUBSTRATE ANALOG COMPLEX FORM.

APO 形式和底物类似复合物形式的脂运载蛋白型前列腺素 D 合酶的结构和动力学分析。

• 发表时间: 2014
• 作者: Shigeru Shimamoto;Tadayasu Ohkubo;Kosuke Aritake;Yoshihiro Urade;Yuji Hidaka
• 通讯作者: Yuji Hidaka