STUDIES ON RATIONAL DESIGN PRINCIPLE FOR FLUORESCENCE ON/OFF SWITCHING

荧光开关合理设计原理的研究

• 批准号: 12470475
• 负责人: NAGANO Tetsuo
• 金额: $ 8.77万
• 依托单位: The 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: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12470475/
• 关键词: fluorescence; probe bioimaging; FRET; photoinduced electron transfer; hydrolase; nitric oxide; zinc ion; reactive oxygenspecies; 活性酸素; ERET

Fluorescence imaging is the most powerful technique currently available for continuous observation of the dynamic intracellular processes of living cells. Fluorescein is widely employed as the core of various fluorescence probes used in imaging important biological effectors, such as nitric oxide and calcium. Despite the extensive use of fluorescein derivatives and the importance of the applications, the mechanism that controls the quantum yield of fluorescence has not been fully established. Without such mechanistic information there can be no rational design strategy to develop new, practically useful fluorescence probes. A plausible mechanism that might control the fluorescence quantum yields of fluorescein derivatives has been suggested to be photoinduced electron transfer from the electron donor moiety to the singlet excited state of the xanthene moiety. Since the fluorescence quantum yields in photoinduced electron transfer reactions may be quantitatively predicted based on the M … More arcus theory of electron transfer, confirmation of the above mechanism would provide a quantitative basis for rational design of fluorescence probes. Further, the stability of fluorescence probes could be increased by accelerating the back electron transfer in the radical ion pair generated by photoinduced electron transfer. However, the occurrence of photoinduced electron transfer in fluorescein-based probes has yet to be established.We obtained the first definitive evidence for the occurrence of photoinduced electron transfer in fluorescein-based probes in which the electron donor moiety is directly linked with the xanthene moiety. Formation of the radical ion pair upon photoirradiation of the fluorescein-based probes was detected by means of laser flash photolysis experiments, which afforded transient absorption spectra showing bands due to the radical cation of the electron donor moiety and the xanthene radical anion. The rates of photoinduced electron transfer and the back electron transfer were determined and analyzed in terms of the Marcus theory of electron transfer. The results provide for the first time a quantitative basis for rational design of fluorescein-based probes with high efficiency in fluorescence ON/OFF switching, as well as the high stability to repeated photoirradiation. We have developed the novel chemical probes for bioimaging of nitric oxide, highly reacrive oxygen species, and zinc ion. Less

荧光成像是目前可用于连续观察活细胞的动态细胞内过程的最强大的技术。黄绿素被广泛用作各种荧光探针的核心，用于成像重要的生物效应物，如一氧化氮和钙。尽管荧光素衍生物的广泛使用和应用的重要性，控制荧光量子产率的机制尚未完全建立。如果没有这样的机制信息，就不可能有合理的设计策略来开发新的，实际上有用的荧光探针。一种可能控制荧光素衍生物的荧光量子产率的合理机制已被认为是从电子供体部分到氧杂蒽部分的单重激发态的光诱导电子转移。由于光诱导电子转移反应中的荧光量子产率可以根据M ...更多信息 电子转移的arcus理论，上述机理的证实将为荧光探针的合理设计提供定量依据。此外，荧光探针的稳定性可以通过加速由光诱导电子转移产生的自由基离子对中的背电子转移来提高。然而，光诱导的电子转移的发生在荧光素为基础的probes尚未建立。我们获得了第一个明确的证据，光诱导的电子转移的发生在荧光素为基础的探针，其中的电子供体部分是直接连接的氧杂蒽部分。通过激光闪光光解实验检测基于荧光素的探针的光照射后的自由基离子对的形成，该实验提供了瞬态吸收光谱，该光谱显示了由于电子供体部分的自由基阳离子和氧杂蒽自由基阴离子的谱带。根据Marcus电子转移理论，测定并分析了光诱导电子转移和背电子转移的速率。结果首次提供了一个定量的基础上，合理设计的荧光素为基础的探针具有高效率的荧光ON/OFF开关，以及高稳定性，重复光照射。我们已经开发了新的化学探针的生物成像一氧化氮，高活性氧物种，和锌离子。少

项目成果

Kenjiro Hanaoka, Kazuya Kikuchi, Yasuteru Urano, Michiko Narazaki, Takashi Yokawa, Shigeru Sakamoto, Kentaro Yamaguchi and Tetsuo Nagano: "Design and synthesis of a novel magnetic resonance imaging contrast agent for selective sensing of zinc ion"Chemistr

Kenjiro Hanaoka、Kazuya Kikuchi、Yasuteru Urano、Michiko Narazaki、Takashi Yokawa、Shigeru Sakamoto、Kentaro Yamaguchi 和 Tetsuo Nagano：“用于选择性感应锌离子的新型磁共振成像造影剂的设计和合成”Chemistr

Kumi Tanaka: "Rational Design of Fluorescein-based Fluorescence Probes.-Mechanism-based Design of a Maximum Fluorescence Probe for Singlet Oxygen-"J.Am.Chem.Soc.. (in press).

Kumi Tanaka：“基于荧光素的荧光探针的合理设计。-单线态氧最大荧光探针的基于机制的设计-”J.Am.Chem.Soc.（出版中）。

Hideo Takakusa, Kazuya Kikuchi, Yasuteru Urano, Hirotatsu Kojima, and Tetsuo Nagano: "A Novel Design Method of Ratiometric Fluorescent Probes Based on Fluorescence Resonance Energy Transfer Switching by Spectral Overlap Integral"Chemistry-A European Journ

Hideo Takakusa、Kazuya Kikuchi、Yasuteru Urano、Hirotatsu Kojima 和 Tetsuo Nagano：“基于光谱重叠积分荧光共振能量转移切换的比率荧光探针的新颖设计方法”Chemistry-A Europe Journ

Sayaka Ueno, Masako Tsukamoto, Tomoya Hirano, Kazuya Kikuchi, Maki K. Yamada, Nobuyoshi Nishiyama, Tetsuo Nagano, Norio Matsuki and Yuji Ikegaya: "Mossy Fiber Zn2+ Spillover Modulates Heterosynaptic N-methyl-D-aspartate Receptor Activity in Hippocampel CA

Sayaka Ueno、Masako Tsukamoto、Tomoya Hirano、Kazuya Kikuchi、Maki K. Yamada、Nobuyoshi Nishiyama、Tetsuo Nagano、Norio Matsuki 和 Yuji Ikegaya：“苔藓纤维 Zn2 溢出调节海马 CA 中的异突触 N-甲基-D-天冬氨酸受体活性

Yasutomo Kawanishi,: "Design and Synthesis of Intramolecular Resonance Energy Transfer Probes for Use in Aqueous Solution"Angewandte Chemie,Int.Ed.. 39. 3438-3440. (2000)

Yasutomo Kawanishi，：“用于水溶液的分子内共振能量转移探针的设计和合成”Angewandte Chemie，Int.Ed.. 39. 3438-3440。