Developing fluorescent probes for the endogenous gaseous transmitters NO and H2S

开发内源性气体递质 NO 和 H2S 荧光探针

• 批准号: 8333986
• 负责人: Michael Pluth
• 金额: $ 24.63万
• 依托单位: UNIVERSITY OF OREGON
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-04-01 至 2014-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8333986
• 关键词: Address; Adsorption; Alzheimer' s Disease; Binding; Binding Sites; Biological; Biological Process; Biology; Cardiovascular system; Catalysis; Cells; Chemistry; Complex; Cytosol; Detection; Development; Dextrans; Doctor of Philosophy; Down Syndrome; Electromagnetics; Electron Transport Complex III; Electronics; Electrons; Endocytosis; Excision; Family; Fluorescence; Fluorescent Probes; Goals; Human body; Huntington Disease; Hydrogen Sulfide; Hydrophobicity; Hypertension; Image; Immune; In Vitro; Individual; Inflammation; Iron; Joints; Laboratories; Lead; Life; Ligands; Measurement; Mentors; Metals; Methodology; Methods; Microscopic; Monitor; Multiple Sclerosis; Natural regeneration; Nerve Degeneration; Nitric Oxide; Nitrogen; Optics; Oxidation-Reduction; Parkinson Disease; Pathway interactions; Penetration; Phase; Photobleaching; Play; Polymers; Positron-Emission Tomography; Postdoctoral Fellow; Process; Production; Property; Reaction; Research; Research Personnel; Resistance; Resolution; Role; Ruthenium; Signal Transduction; Stimulus; Time; Tissues; Transition Elements; United States National Institutes of Health; Vasodilation; Water; Work; abstracting; base; blood pressure regulation; carcinogenesis; design; dextran; extracellular; fluorophore; functional group; in vivo; interest; metal complex; molecular recognition; nervous system disorder; physical property; prevent; scaffold; sensor; single walled carbon nanotube; small molecule; tool; tumor growth

Project Summary/Abstract The candidate received his Ph.D. from the UC Berkeley, under the joint direction of Kenneth Raymond and Robert Bergman where he studied host-guest chemistry, molecular recognition, and catalysis in water-soluble supramolecular complexes. He is currently an NIH postdoctoral fellow in Stephen Lippard's laboratory at MIT working on developing fluorescent probes for nitric oxide. The candidate's research interests span the field of molecular recognition with a specific focus on how microscopic processes lead to the recognition of individual atoms, functional groups and molecules. The candidate will use his background in mechanistic studies and molecular recognition to pursue his research interests as a principle investigator. His independent research will focus on the development of new tools for the detection and imaging of small molecules in biology. Nitric oxide (NO) and hydrogen sulfide (H2S) are now accepted as biologically important gaseous transmitters. Both NO and H2S are produced endogenously and are finely regulated by the body. Nitric oxide is beneficial for vasodilation and immune activity at low cellular concentrations but overproduction can lead to the proliferation of reactive NO species that have been implicated in carcinogenesis and several degenerative neurological disorders, including Alzheimer's (AD), Parkinson's, and Huntington's disease, as well as multiple sclerosis. Similarly, H2S has been implicated in AD, Downs syndrome and other forms of metal deficiency. H2S also plays an active role in inflammation and in blood pressure regulation. Despite the recognized importance of both of these gaseous transmitters, the current methods for detection in live cells are limited. The postdoctoral phase of the proposed research will focus on the development of new NO-selective fluorescent probes that address current limitations of NO detection. Transition metal based NO binding sites will be used to develop probes that can reversibly bind NO and probes that emit in the NIR. The proposed family of fluorescent probes will use paramagnetic (S=1/2) metals serving the dual role as both fluorescence quencher and NO binding site. Coordination of NO will form a diamagnetic (S=0) complex and restore the fluorescence of the pendant fluorophore. Adsorption or covalent attachment of such complexes to solubilized single-walled carbon nanotubes (SWNTs) will be used to develop NO-selective probes that emit in the NIR. The independent research phase of the proposed research will investigate the design of H2S-selective fluorescent probes for the imaging of endogenously produced H2S. Currently, such H2S detection methods are lacking and most measurements rely on bulk tissue measurements. The new H2S-selective fluorescent probes for use in live cells will provide much needed tools for the study of the biological functions of H2S. The unique physical properties of H2S will all be exploited in the design of H2S-selective fluorescent probes. Fluorophores will be derivatized with specially designed protecting groups that can only be removed by H2S. Removal of the fluorophore protecting group will restore the fluorescence, thus forming a turn-on probe for H2S.

项目摘要/摘要 候选人获得了博士学位。从加州大学伯克利分校，在肯尼斯·雷蒙德的联合指导下 罗伯特·伯格曼（Robert Bergman 超分子复合物。他目前是MIT的Stephen Lippard实验室的NIH博士后研究员 致力于开发一氧化氮的荧光探针。候选人的研究兴趣跨越了 分子识别，特别关注微观过程如何导致个人识别 原子，官能团和分子。候选人将利用他的机械研究背景和 分子认可作为主要研究者的研究兴趣。他的独立研究将 专注于开发新工具，用于检测和成像生物学中的小分子。 一氧化氮（NO）和硫化氢（H2S）现在被接受为生物学上重要的气态 发射器。 NO和H2均可内源产生，并由人体细节调节。一氧化氮是 在低细胞浓度下有益于血管舒张和免疫活性，但产生过量会导致 反应性无物种的增殖，与癌变和几种退化有关 神经系统疾病，包括阿尔茨海默氏症（AD），帕金森氏症和亨廷顿氏病，以及多个 硬化。同样，H2S与AD，Downs综合征和其他形式的金属缺乏有关。 H2S 在炎症和血压调节中也起着积极作用。尽管很重要 在这两个气态发射器中，现有的现场检测方法受到限制。 拟议研究的博士后阶段将重点介绍新的无选择的发展 荧光探针解决了无检测的当前局限性。过渡金属无结合位点 将用于开发可以可逆地绑定NO的探针和在NIR中发出的探针。提议 荧光探针家族将使用顺磁性（S = 1/2）金属，均为双重作用 淬灭和无绑定位点。 NO的协调将形成Diamagnetic（S = 0）复合物并恢复 吊坠荧光团的荧光。这种复合物的吸附或共价附着在溶解化 单壁碳纳米管（SWNT）将用于开发NIR中发出的无选择探针。 拟议研究的独立研究阶段将研究H2S选择性的设计 内源产生的H2s成像的荧光探针。当前，此类H2S检测方法是 缺乏和大多数测量取决于散装组织测量。新的H2S选择性荧光探针 用于实时细胞将提供急需的工具来研究H2S的生物学功能。独特 H2S的物理特性都将在H2S选择性荧光探针的设计中被利用。荧光团 将通过专门设计的保护组衍生，只能由H2S删除。去除 荧光团保护组将恢复荧光，从而形成对H2S的转交探针。