Models of the ctiue sites of metalloproteins : synthetic, structural, and functional studies on metallopeptides

金属蛋白的结构位点模型：金属肽的合成、结构和功能研究

• 批准号: 08454213
• 负责人: YAMAMURA Takeshi
• 金额: $ 4.93万
• 依托单位: Science University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1996
• 资助国家: 日本
• 起止时间: 1996 至 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08454213/
• 关键词: Zinc finger; Cys-X_2-Cys; Zn^<2+>, M^<2+>; hydrolases; His-Glu-X_2-His; Zn^<2+>; molecular dynamics; molecular orbital method; artificial protein; multiple porphyrin; His-Gm-X_2-His; マルチポルフィソン系; ペプチド; 亜鉛錯体; リューマチ; プロテインキナーゼC; ポテンショメトリー; 金

1) We prepared the mercury peptide, [(Boc-Cys-Pro-Leu-Cys-Gly-Ala-OMe)Hg], and studied the coordination and conformation structures by EXAFS, ROESY, distance geometry (DG), restrained molecular dynamics (RMD) calculations, -. Comparison of the structure with that of [(Boc-Cys-Pro-Leu-Cys-OMe)Hg] revealed that both of these have a mirror type gamma-tum structure.2) We clarified the structure of [(Boc-Cys-Pro-Leu-Cys-OMe)(S-tert-C_4H_9)Hg] by the same way as above. This study clarified that the conformation of Cys-X_2-Cys does not depend on the kinds of metal ions, but depends on the S-M-S bite angle, and the peptide conformation obtained for this compound agreed with that of the zinc binding sites of zinc finger motifs3)We synthesized [(Boc-Cys-Pro-Leu-Cys-OMe)Zn] and [(Boc-Cys-Pro-Leu-Cys-Gly-Ala-OMe)Zn] in terms of zinc finger motifs. We studied the substitution reactions of the zinc sites of these compounds. The structure of the former was discussed by molecular orbital calculations, MD calculations, EXAFS, NMR.4) We studied the complex formation between Boc-Glu-Thr-Ile-His-OMe and Zn^<2+> with respect to the zinc binding sites of proteases. The structural study by ROESY, RMD and MO suggested that this system generates a species which is structurally similar to the corresponding sites of proteases.5) We de novo designed and synthesized an artificial protein, which is capable of incorporating multiple porphyrins into its His sites, on the basis of template assisted synthetic protein approach, where a picket fence porphyrin was adopted as the template. This peptide porphyrin incorporated another types of porphyrins into the His sites forming a series of trimeric aggregates, thus achieved a 12 hetero-porphyrin system.

1)我们制备了汞肽[(Boc-Cys-Pro-Leu-Cys-Gly-Ala-OMe)Hg]，并通过EXAFS、ROESY、距离几何(DG)、约束分子动力学(RMD)计算研究了配位和构象结构，-。与[(Boc-Cys-Pro-Leu-Cys-OMe)Hg]的结构比较表明，两者都具有镜像型γ-tum结构。2)我们通过与上述相同的方式阐明了[(Boc-Cys-Pro-Leu-Cys-OMe)(S-tert-C_4H_9)Hg]的结构。本研究阐明Cys-X_2-Cys的构象并不取决于金属离子的种类，而是取决于S-M-S咬合角，并且该化合物获得的肽构象与锌指基序的锌结合位点一致3)我们合成了[(Boc-Cys-Pro-Leu-Cys-OMe)Zn]和 [(Boc-Cys-Pro-Leu-Cys-Gly-Ala-OMe)Zn] 就锌指基序而言。我们研究了这些化合物的锌位点的取代反应。通过分子轨道计算、MD计算、EXAFS、NMR对前者的结构进行了讨论。4)我们研究了Boc-Glu-Thr-Ile-His-OMe与Zn^<2+>在蛋白酶的锌结合位点上形成的复合物。 ROESY、RMD和MO的结构研究表明，该系统产生了一个在结构上与蛋白酶相应位点相似的物种。5)我们在模板辅助合成蛋白方法的基础上，以栅栏卟啉为模板，从头设计并合成了一种能够将多个卟啉掺入其His位点的人工蛋白。该肽卟啉将另一种类型的卟啉掺入到His位点中，形成一系列三聚体聚集体，从而实现了12杂卟啉系统。

项目成果

Hiroteru Nishikawa, Masato Ushiyama, Takeshi Yamamura: "De Novo Designed Heme-binding Peptide-porphyrins. Excitonic Analyses of the UV-vis and CD Spectra" Peptide Science. (in press.). (1999)

Hiroteru Nishikawa、Masato Ushiyama、Takeshi Yamamura：“从头设计血红素结合肽卟啉。紫外可见光谱和 CD 光谱的激子分析”肽科学。

Masato Ushiyama, Hiroteru Nishikawa, Yuka Morio, Takeshi Yamamura: "Construction of Multi Porphyrin Systems Using Histidine Containing 4-Helix Bundle Porphyrin and Heme" Peptide Science. (in press.). (1999)

Masato Ushiyama、Hiroteru Nishikawa、Yuka Morio、Takeshi Yamamura：“使用含有 4 螺旋束卟啉和血红素的组氨酸构建多卟啉系统”肽科学。

Hiroteru Nishikawa,: "De Nouo Designed Heme-Bincling Peptide Porphyrins.Excltonic Analyses of the UV-vis and CD Spectina" Peptide Science. in press. (1999)

Hiroteru Nishikawa，：“De Nouo 设计的血红素结合肽卟啉。UV-vis 和 CD Spectina 的激子分析”肽科学。

Takeshi Yamamura, Masaya Arai, Tsutomu Yamane, Takeshi Ukai, Masato Ushiyama, and Hiroshi Hirota: "Conformation Control of Model Peptides by Metal Ions. A New Type of Turn Structure Found in [(Boc-Cys-Pro-Leu-Cys-Gly-Ala-OMe)Hg]" Bull.Chem.Soc.Jpn.69. 222

Takeshi Yamamura、Masaya Arai、Tsutomu Yamane、Takeshi Ukai、Masato Ushiyama 和 Hiroshi Hirota：“金属离子对模型肽的构象控制。在 [(Boc-Cys-Pro-Leu-Cys-Gly) 中发现的新型转角结构

Masato Ushiyama, Atsushi Yoshino, Takeshi Yamamura, Yasuo Shida, Fumio Arisaka: "A 12-Porphyrin System : Syntheses of Peptide Porphyrins with Multiple Histidines and the Aggregation Behavior in the Presence of Hemin" Bull.Chem.Soc.Jpn.72 (in press.). (199

Masato Ushiyama、Atsushi Yoshino、Takeshi Yamamura、Yasuo Shida、Fumio Arisaka：“12-卟啉系统：具有多个组氨酸的肽卟啉的合成以及氯化血红素存在下的聚集行为”Bull.Chem.Soc.Jpn.72（in