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分子的磷酸化和去磷酸化来调节信号转导。激酶和磷酸酶活性的检测技术对于阐明信号转导机制至关重要。荧光比率成像是一种灵敏、非破坏性和准确的方法，用于检测活细胞内的这种酶反应。在本研究中，我们设计了一种荧光探针，它可以特异性地与磷酸酶反应，改变磷酸酶的荧光特性。（1）探针应该具有开关功能，通过该开关功能，探针的激发光谱在与磷酸酶反应时改变，以及（2）探针中的反应位点的结构可以容易地改变，这样探针就能特异性地与另一种磷酸酶反应，而这种磷酸酶能识别不稳定位点的独特结构。首先，我们选择7-羟基香豆素作为 ...更多信息 探针的基本结构。重要的是，我们的初步实验表明，羟基的pK_a是由空间位置接近羟基的阴离子基团控制的，并影响化合物的激发光谱。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，使得磷酸酶去除磷酸基团导致羟基的pKa改变，从而导致探针的激发光谱改变。此外，脂肪族连接基允许修饰接近磷酸基团的结构。结果表明，该探针能与酸性磷酸酶发生特异性反应，改变了酸性磷酸酶的激发光谱。此外，激发光谱的变化大到足以进行比率测量。这些结果为荧光比率探针的设计提供了有价值的信息，我们的策略在生物成像领域具有很大的应用潜力。少

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

Inhibition of Presynaptic Activity by Zinc Released From Mossy Terminals During Tetanic Stimulation.

强直刺激期间苔藓末梢释放的锌对突触前活动的抑制。

• 发表时间: 2006
• 期刊: J.Neurosci.Res. 83
• 作者: A.Minami;N.Sakurada;S.Fuke;K.Kikuchi;T.Nagano;N.Oku;A.Takeda
• 通讯作者: A.Takeda

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