The Reaction Mechanism and Subsites of RNase T_2 family

RNase T_2家族的反应机制及亚位点

• 批准号: 08680663
• 负责人: NAKAMURA Kazuo
• 金额: $ 1.54万
• 依托单位: Showa University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1996
• 资助国家: 日本
• 起止时间: 1996 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08680663/
• 关键词: Ribonuclease; X-ray analysis; Enzyme-substrate complex; X線構造解析; 基質複合体; サブサイト

During the present study we have successfully determined the two crucial crystal structures of RNase RNAP-Rh, i.e., RNase RNAP-Rh+d (ApG) complex and RNAP-Rh (Y57W) +d (ApC) complex. Diffraction data of the crystals were collected using an oscillation camera (R-AXISIIc manufactured by Rigaku). The crystal structures were determined based on the atomic coordinates of RNase Rh, and refined using the program X-PLOR.The refinements were done successfully, and the R-values of both complexes were coverged to under 0.20. (1) we found out that there are two hydrogen-bonds between the adenine base and the enzyme, and three ones between the phosphate group and the enzyme. (2) We also found out that the subsite B2 consists of Gln32, Pro92, Ser93, Asn94, Gln95 and Phe101.

在本研究中，我们成功确定了 RNase RNAP-Rh 的两个关键晶体结构，即 RNase RNAP-Rh+d (ApG) 复合物和 RNAP-Rh (Y57W) +d (ApC) 复合物。使用振荡相机(Rigaku制造的R-AXISIIc)收集晶体的衍射数据。根据RNase Rh的原子坐标确定了晶体结构，并使用X-PLOR程序进行了精修。精修成功完成，两种配合物的R值均覆盖到0.20以下。 (1)我们发现腺嘌呤碱基与酶之间存在两个氢键，磷酸基与酶之间存在三个氢键。 (2)我们还发现B2亚位点由Gln32、Pro92、Ser93、Asn94、Gln95和Phe101组成。

项目成果

桐野豊,中村和郎他: "基礎薬学 物理化学(4月発刊予定)" 広川書店, 300 (1997)

Yutaka Kirino、Kazuo Nakamura 等：《基础药剂学物理化学（预定 4 月出版）》广川书店，300（1997）

S.Parry et al.: "Structural Analysis & Molecular Model of a self-Incompatibility RNase from Wild Tomato" Plant Physiol.Vol.116. 463-469 (1998)

S.Parry 等人：“结构分析

S. Parry et al.: "Structural Analysis & Molecular Model of a Self-Incom-patibility RNase from Wild Tomato" Plant Physiol.116. 463-469 (1998)

S. Parry 等人：“结构分析