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.

项目总结/摘要 候选人获得了博士学位。来自加州大学伯克利分校，在肯尼斯·雷蒙德的联合指导下， 罗伯特伯格曼在那里他研究主客体化学，分子识别，和催化在水溶性 超分子复合物他目前是麻省理工学院斯蒂芬·利帕德实验室的NIH博士后研究员 研究一氧化氮的荧光探针候选人的研究兴趣涵盖以下领域： 分子识别，特别关注微观过程如何导致个体识别 原子、官能团和分子。候选人将利用他在机械研究方面的背景， 分子识别，以追求他的研究兴趣作为一个主要的研究者。他的独立研究将 专注于开发用于生物学中小分子检测和成像的新工具。 一氧化氮（NO）和硫化氢（H2S）是目前公认的重要的生物气体 发射器NO和H2S都是内源性产生的，并受到身体的精细调节。氧化氮是 在低细胞浓度下有益于血管舒张和免疫活性，但过量生产可导致 增殖的活性NO物种已牵连在致癌作用和几个退行性变 神经系统疾病，包括阿尔茨海默病（AD）、帕金森病和亨廷顿病，以及多种神经系统疾病， 硬化症类似地，H2S与AD、唐斯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积极作用。尽管公认的重要性 对于这两种气体递质，目前在活细胞中检测的方法是有限的。 拟议研究的博士后阶段将集中在开发新的NO选择性 荧光探针解决了目前NO检测的局限性。基于过渡金属的NO结合位点 将被用于开发探针，可以可逆地结合NO和探针，在近红外发射。拟议 荧光探针家族将使用顺磁性（S=1/2）金属，其充当荧光探针和非荧光探针的双重作用。 猝灭剂和NO结合位点。NO的配位将形成抗磁性（S=0）络合物，并恢复铁磁性。 悬垂荧光团的荧光。这种复合物吸附或共价连接到溶解的 单壁碳纳米管（SWNT）将用于开发在NIR中发射的NO选择性探针。 拟议研究的独立研究阶段将调查H2S选择性 荧光探针用于内源性产生的H2S的成像。目前，这样的H2S检测方法是 缺乏且大多数测量依赖于大块组织测量。新型H2S选择性荧光探针 将为研究H2S的生物学功能提供急需的工具。独特的 H2S的物理性质都将在H2S选择性荧光探针的设计中被利用。荧光团 将用专门设计的保护基团衍生化，这些保护基团只能被H2S除去。移除 荧光团保护基团将恢复荧光，从而形成H2S的开启探针。