Specific Metal Ion Recognition Mechanism by Protein Molecule

蛋白质分子识别特定金属离子的机制

• 批准号: 13640553
• 负责人: KOHZUMA Takamitsu
• 金额: $ 2.3万
• 依托单位: Ibaraki University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13640553/
• 关键词: Metalloprotein; Copper Protein; Molecullar Recognition; Pseudoazurin; Plastocyanin; UV Resonance Raman; 金属蛋白質; 弱い相互作用; ブルー銅蛋白質; スタッキング; 常磁性NMR; 紫外共鳴ラマンスペクトル; 銅蛋白質

The UV resonance Raman spectrum of pseudoazurin gives a copper coordinated histidine imidazole band at 1385 cm^<-1>. A characteristic histidine imidazolium Raman band at 1408 cm^<-1> is detected in the UVRR spectrum of Cu(I) pseudoazurin at acidic pH. This supports the protonation of the His81 ligand in the reduced protein at low pH values. The imidazolium Pseudoazurin substituted with Zn^<2+> gave characteristic Raman Bands for the metal ion coordinated imidazole at 1388 cm^<-1> at neutral pH. The histidine imidazole band diminishes in intensity in Zn^<2+> pseudoazurin upon lowering the pH. In all of the metal-substituted pseudoazurins an imidazolium band at 1408 cm^<-1> appears at pH < 4.0. Pseudoazurin substituted with Zn^<2+>, Cd^<2+>, and Ni^<2+> give very similar pH dependence of their X-Pro Raman bands as apo-pseudoazurin. These findings indicate that the metal ion is released from the active site of Zn^<2+>, Cd^<2+>, and Ni^<2+> pseudoazurin at acidic pH accompanied by a change in the protein's secondary structure. This behavior is clearly absent in both the Cu(I) and CU(II) proteins indicating that pseudoazurin discriminates between copper and the other metal ions studied.

假天青的UV共振拉曼光谱在1385 cm-1处给出铜配位的组氨酸咪唑带<-1>。在<-1>酸性pH下，在Cu（I）假天青的UVRR光谱中检测到1408 cm-1处的特征性组氨酸咪唑拉曼带。这支持低pH值下还原蛋白质中His 81配体的质子化。用Zn^&lt;2+&gt;取代的咪唑类假天青在中性pH下在1388 cm ↑ [2]处给出了金属离子配位咪唑的特征拉曼谱带<-1>。在Zn^&lt;2+&gt;的假天青中，组氨酸咪唑谱带的强度随pH值的降低而减弱。在所有的金属取代的假天青中，在pH &lt; 4.0时出现了1408 cm ↑ [2]处的咪唑类<-1>谱带。用Zn^&lt;2+&gt;、Cd^&lt;2+&gt;和Ni^&lt;2+&gt;取代的假天青的X-Pro拉曼谱带与脱辅基假天青的pH依赖性非常相似。这些发现表明，在酸性pH条件下，Zn^&lt;2+&gt;、Cd^&lt;2+&gt;和Ni^&lt;2+&gt;假天青蛋白的活性部位释放出金属离子，并伴随着蛋白质二级结构的变化。这种行为在Cu（I）和Cu（II）蛋白中明显不存在，表明假天青蛋白区分铜和所研究的其他金属离子。

项目成果

Hyato Morita: "Heteronuclear NMR and UV-Resonance Raman Studies on the molecular mechanism of Zn^<2+> sensing with cyanobacterial transcription factor, SmtB"J.Inorg.Biochem.. 96. 196 (2003)

Hyato Morita：“利用蓝藻转录因子 SmtB 检测 Zn^2> 分子机制的异核 NMR 和 UV 共振拉曼研究”J.Inorg.Biochem.. 96. 196 (2003)

Claudio Fernandez: "Paramagnetic ^1H NMR study of the metal site in Cu(II) and Co(II)-pseudoazurin"J.Inorg.Biochem.. 86. 217 (2001)

Claudio Fernandez：“Cu(II) 和 Co(II)-pseudoazurin 中金属位点的顺磁 ^1H NMR 研究”J.Inorg.Biochem.. 86. 217 (2001)

Hiroko Kodama: "Biochemical Indicator for Evaluation of Connective Tissue Abnormalities in Menkes' Disease"The Journal of Pediatics. 726-728 (2003)

Hiroko Kodama：“评估门克斯病结缔组织异常的生化指标”儿科杂志。

Katsuko Sato: "UV Resonance Raman and NMR Spectroscopic Studies on the pH Dependent Metal Ion Release from Pseudoazurin"Inorg.Chim.Acta. 339. 383-392 (2002)

Katsuko Sato：“Pseudoazurin 中 pH 依赖性金属离子释放的紫外共振拉曼和核磁共振光谱研究”Inorg.Chim.Acta。

Christopher Dennison: "Unusual Properties of Plastocyanin from the Dryopteris crassirhizoma"Biochemistry. 41. 552-560 (2002)

克里斯托弗·丹尼森（Christopher Dennison）：“来自粗鳞毛蕨的质体蓝素的不寻常特性”生物化学。