CRYSTAL STRUCT. OF CYCLIC NUCLEOTIDE PHOSPHODIESTERASE

晶体结构。

• 批准号: 6386586
• 负责人: Hengming Ke
• 金额: $ 18.17万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-04-01 至 2004-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6386586
• 关键词: 3'5' cyclic nucleotide phosphodiesterase; X ray crystallography; active sites; crystallization; cyclic AMP; cyclic GMP; drug design /synthesis /production; enzyme structure; enzyme substrate complex; isozymes; phosphodiesterase inhibitors; physical model

DESCRIPTION: (Verbatim from the Applicant's Abstract) Cyclic nucleotide phosphodiesterase (PDE) catalyzes the hydrolysis of adenosine 3',5'-cyclic monophosphate (cAMP) and guanosine 3',5'-cyclic monophosphate (cGMP) to produce, respectively, 5'-AMP and 5'-GMP. PDE is a key enzyme to control cellular concentrations of cAMP that is known as "second messenger" and mediates the response of cells to a wide variety of hormones and neurotransmitters. Ten families and twenty two subtypes of human PDE have been identified. The mRNAs of the 22 subtype PDEs are further spliced to generate over 60 isoforms of PDE. The distinct isoforms of PDE are located in different cellular compartments and possess different specificity of substrate. These two features of PDE have attracted great attention from pharmaceutical companies in the past decade. Many selective PDE inhibitors have been studied as therapeutic agents such as cardiotonic agents, vasodilators, antiasthma, atithrombic compounds, smooth muscle relaxants and antidepressants. For example, VIAGRA, an inhibitor of PDE5, is a prescription drug for erectile dysfunction of male patients. This proposal aims at characterization of substrate specificity and inhibitor selectivity by the approach of crystallography. The specific aims are to determine crystal structures of PDEs and their complexes with inhibitors including (1) the catalytic domain of PDE4B, (2) the catalytic domain of PDE4B complexed with the inhibitors rolipram and iodonated cAMP, (3) full length PDE4D and its complexes with the inhibitors rolipram and denbufylline and (4) the catalytic domain of PDE3 and its complex with cilostamide. The structures in this proposal will reveal the details of inhibitor binding at the active site and provide insight into catalytic mechanism. Docking cGMP into the active site of PDE4B, together with the structure of PDE4B-cAMP analog, will shed light on the substrate specificity. Comparison of the structures of PDE-inhibited complexes will shed light on the selectivity of inhibition of different families of PDE, and thus provide a structural basis for design of selective drugs. The structures will be determined by multiple isomorphous replacement, multiwavelength anomalous diffraction, or molecular replacement. The structural models will be built with the program O and refined by the program CNS.

描述：（逐字摘自申请人摘要）环核苷酸 磷酸二酯酶 (PDE) 催化 3',5'-环腺苷的水解 单磷酸盐 (cAMP) 和鸟苷 3',5'-环单磷酸盐 (cGMP) 分别产生5'-AMP和5'-GMP。 PDE是控制的关键酶 被称为“第二信使”的 cAMP 的细胞浓度 介导细胞对多种激素的反应 神经递质。人类 PDE 的 10 个家族和 22 个亚型已被 确定。 22个亚型PDE的mRNA进一步剪接生成 超过 60 种 PDE 亚型。 PDE 的不同异构体位于不同的位置 细胞区室并具有不同的底物特异性。这两个 PDE的特点引起了制药企业的高度关注 过去十年。许多选择性 PDE 抑制剂已被研究作为治疗药物 强心剂、血管扩张剂、抗哮喘剂、抗血栓剂等药剂 化合物、平滑肌松弛剂和抗抑郁剂。例如，伟哥 (VIAGRA) PDE5抑制剂，是治疗男性勃起功能障碍的处方药 患者。该提案旨在表征底物特异性和 通过晶体学方法进行抑制剂选择性。具体目标是 确定 PDE 及其与抑制剂的复合物的晶体结构 包括(1)PDE4B的催化结构域，(2)PDE4B的催化结构域 与抑制剂咯利普兰和碘化 cAMP 复合，(3) 全长 PDE4D 及其与抑制剂咯利普兰和登布茶碱的复合物以及 (4) PDE3 的催化结构域及其与西洛酰胺的复合物。结构 在该提案中将揭示抑制剂在活性位点结合的细节 位点并提供对催化机制的深入了解。将 cGMP 对接至活性物质 PDE4B 位点以及 PDE4B-cAMP 类似物的结构将脱落 光对底物的特异性。结构比较 PDE 抑制复合物将揭示 PDE 抑制的选择性 不同族的偏微分方程，从而为设计提供结构基础 选择性药物。结构将由多个同晶决定 置换、多波长异常衍射或分子置换。 结构模型将使用 O 程序构建并通过 中枢神经系统程序。