Design and synthesis of visualization sensor molecules for bio-imaging

用于生物成像的可视化传感器分子的设计与合成

• 批准号: 18310144
• 负责人: KIKUCHI Kazuya
• 金额: $ 11.56万
• 依托单位: Osaka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2006
• 资助国家: 日本
• 起止时间: 2006 至 2007
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18310144/
• 关键词: fluorescence ratio imaging; fluorescent probe; phosphatase; excitation spectrum; signal transduction; 希土類蛍光プローブ; 時間分解スペクトル; calpain; MRI; 蛍光センサー; 蛋白質相互作用; 長寿命蛍光; 蛍光顕微鏡; イメージインテンシファイア; 分子デザイン; 可視化解析

Kinases and phosphatases regulate signal transduction through phosphorylation and dephosphorylation of various biological molecules. The techniques for detecting the activity of kinases and phosphatases are essential to the elucidation of signal transduction mechanism. The fluorescence ratio imaging is a sensitive, undestructive and accurate method for the detection of such enzymatic reactions inside living calls. In this research, we created fluorescent probes, which specifically react with phosphatase and change its fluorescence properties.Our design strategy is based on the following requirements; (1) The probes should have a switch function, by which the excitation spectrum of the probe changes upon the reaction with phosphatase, and (2) the structure of the reactive site in the probe can be easily modified, so that the probe specifically reacts with a different kind of phosphatase, which recognizes a distinctive structure of the restive site. First, we chose 7-hydr xycoumarin as … More a basic structure of the probe. Importantly, our preliminary experiments show that the pK_a of the hydroxyl group is controlled by the anionic group sterically close to the hydroxyl group and affects excitation spectrum of the compound. We, therefore, introduced phosphate group into the proximal position of the hydroxyl group via the aliphatic ester linkage, so that removal of the phosphate group by phosphatase results in the alteration of pKa of the hydroxyl group, leading to the change of the excitation spectrum of the probe. In addition, the aliphatic linker allows modification of the structure close to the phosphate group. It was demonstrated that the probe specifically reacted with acid phosphatase and changed its excitation spectrum. Furthermore, the change of the excitation spectrum was large enough to conduct ratio measurements. These results provides valuable information about design of fluorescent ratio probes and our strategy has great potential toward application in the field of bio-imaging. Less

激酶和磷酸酶通过各种生物学分子的磷酸化和去磷酸化来调节信号转移。检测激酶和磷酸酶活性的技术对于阐明信号转移机制至关重要。荧光比成像是一种敏感，非结构性和准确的方法，用于检测活呼叫中这种酶促反应。在这项研究中，我们创建了荧光问题，该问题特别与磷酸酶反应并改变其荧光性能。我们的设计策略基于以下要求； （1）探针应具有开关函数，探针的兴奋光谱在与磷酸酶反应时发生变化，并且（2）探针中反应性位点的结构可以很容易地修饰，因此探针与另一种类型的磷酸酶特异性反应，从而识别出可休息位点的独特结构。首先，我们选择7-HYDR Xycoumarin作为……更多的探针基本结构。重要的是，我们的初步实验表明，羟基的PK_A受到靠近羟基的阴离子基团的控制，并影响该化合物的兴奋光谱。因此，我们通过脂肪族酯键将磷酸基团引入羟基的近端位置，以便通过磷酸酶去除磷酸基团导致羟基PKA改变羟基的PKA，从而导致探针激发谱的变化。此外，脂肪族接头允许修改靠近磷酸盐基团的结构。证明探针与酸性磷酸酶特异性反应并改变了其兴奋光谱。此外，兴奋光谱的变化足以进行比率测量。这些结果提供了有关荧光比问题设计的有价值的信息，我们的策略具有在生物成像领域应用的巨大潜力。较少的

项目成果

Design and synthesis of an enzyme activity-based labeling molecule with fluorescence spectral change

• DOI: 10.1021/ja0657307
• 发表时间: 2006-12-20
• 期刊: JOURNAL OF THE AMERICAN CHEMICAL SOCIETY
• 影响因子: 15
• 作者: Komatsu, Toru;Kikuchi, Kazuya;Nagano, Tetsuo
• 通讯作者: Nagano, Tetsuo

A Gd^<3+>-Based Magnetic Resonance Imaging Contrast Agent Sensitive to beta-Galactosidase Activity Utilizing a Receptor-Induced Magnetization Enhancement(RIME)phenomenon

利用受体诱导磁化增强(RIME)现象的对β-半乳糖苷酶活性敏感的基于Gd^3>的磁共振成像造影剂

• 发表时间: 2008
• 期刊: Chemistry-A European Journal 14
• 作者: Shin Mizukami;Kenjiro Hanaoka
• 通讯作者: Kenjiro Hanaoka

Modulation of luminescence intensity of lanthanide complexes by photoinduced electron transfer and its application to a long-lived protease probe

• DOI: 10.1021/ja060729t
• 发表时间: 2006-05-31
• 期刊: JOURNAL OF THE AMERICAN CHEMICAL SOCIETY
• 影响因子: 15
• 作者: Terai, Takuya;Kikuchi, Kazuya;Nagano, Tetsuo
• 通讯作者: Nagano, Tetsuo

Selective photoinactivation of protein function through environment-sensitive switching of singlet oxygen generation by photosensitizer

• DOI: 10.1073/pnas.0611717105
• 发表时间: 2008-01
• 期刊: Proceedings of the National Academy of Sciences
• 作者: T. Yogo;Y. Urano;A. Mizushima;Hisato Sunahara;Takanari Inoue;K. Hirose;M. Iino;K. Kikuchi

Toward Bifunctional Antibody Catalysis

迈向双功能抗体催化

• 发表时间: 2006
• 期刊: Bioorg. Med. Chem. 14
• 作者: K.Kikuchi;R.B.Hannak;M.J.Guo;A.J.Kirby;D.Hilvert
• 通讯作者: D.Hilvert