NEW PROBES AND REAGENTS FOR BIOLOGICAL STUDIES

用于生物学研究的新探针和试剂

• 批准号: 3274555
• 负责人: JOHN F KEANA
• 金额: $ 16.86万
• 依托单位: UNIVERSITY OF OREGON
• 依托单位国家: 美国
• 财政年份: 1980
• 资助国家: 美国
• 起止时间: 1980-02-01 至 1993-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3274555
• 关键词: Diels Alder reaction; adenosine triphosphate; adenosinetriphosphatase; amines; anthracenes; cell membrane; chemical synthesis; contrast media; drug metabolism; electron microscopy; electron spin resonance spectroscopy; ligands; liposomes; magnetic resonance imaging; membrane activity; membrane proteins; neutron radiation; nitrogen; nitrous oxide; phosphatidylinositols; phosphonate; radionuclide therapy; reagent /indicator; receptor; singlet oxygen; stable isotope

The central theme of the overqall research program is the design, chemical synthesis, characterization, and collaborative application of new probes and reagents designed to overcome some of the currently serious obsticals in biological and medical research. Four subareas form the focus of this five year project. a) New Contrast-Enhancing Agents for Magnetic Resonance Imaging. The long-term objective is to develop improved, clinically acceptable paramagnetic agents that selectively enhance the contrast of targety tissues and organs in MRI appplications. In order to maximize effectiveness of the contrast agent, moleccules to which a larage number of paramagnetic centers may be attached in a chemically well defined manner will be synthesized. The "molecular amplifier" is then attached to the targeting device, e. g., an antibody or liposome, through a chemically reactive arm. Thus, one might be able toachieve excellent contrast with a relatively small number of paramagnetic centers. The amplifier concept is expected to be applicable to other areas, e. g., boron-10 neutron capture therapy. Amplification wil also be sought through the synthesis of highly paramagnetic liposomes and through modification of the "molecular box", apoferritin. b) New ESR Probes for Singlet Oxygen. Singlet ocygen is a likely oxidant associated with the photodynamic effect in biological systems. A water soluble analog of our successful anthracene nitroxide-based ESR singlet oxygen probe is needed and this is our objective. The significance of thes segment is the development of new tools for the study of oxidation, a fundamental process in biological systems. c) New Small, Highly Electron Dense Electron Microscopy Labels. Methods will be developed for the introduction of a radiolabel in our new heteropolytungstate (HPT)-based EM labels, permitting independent quantitation of labeling levels in a biological system. We intend to synthesize a photolabeled, HPT-labeled ATP for a collaborative study aimed at determining the location of the ATP binding sites in F0F1-ATPase, a ubiquitous ATP-synthesizing enzyme. d) Synthesis and Applications of Perdeutero N-Proxyl Nitroxide Spin Labels. With the aim of enhancing the sensitivity of the chemically robust proxyl nitroxide spin labels developed in our laboratory, we intend to develop a convenient synthetic route to perdeutero-15N-proxyls. A chemically well defined spin labeled phosphatidylinositol (PI) as well as a labeled posphonate analog will be synthesized for collaborative biophysical investigation of lipid-protine interactions occurring between PI and membrane proteins associated with membrane recptor transduction.

Overqall研究计划的中心主题是设计， 化学合成、表征和协同应用 一系列新的探针和试剂，旨在克服一些 目前在生物和医学研究中有严重的迟疑。 四个分领域构成了这一五年计划的重点。A)新增 磁共振成像的对比度增强剂。这个 长期目标是开发出改进的、临床可接受的 具有选择性增强对比度的顺磁剂 磁共振成像应用中的组织器官损害。为了 最大限度地提高造影剂的有效性，对其分子 大量的顺磁中心可能附着在 化学上定义明确的方式将被合成。“分子” 然后将“放大器”连接到目标设备上，例如 抗体或脂质体，通过化学反应的手臂。因此，一个 可能能够在相对较小的情况下实现良好的对比度 顺磁中心的数量。放大器的概念是预期的 适用于其他领域，例如，硼-10中子捕获 心理治疗。还将通过合成来寻求放大 并通过对高顺磁性脂质体的修饰 “分子盒”，即脱铁蛋白。B)新的单线态ESR探头 氧气。单线态氧很可能是一种与 生物系统中的光动力效应。一种水溶性类似物 我们成功的基于氧化菲的电子自旋共振单线态氧 探测是必要的，这是我们的目标。的重要意义。 这一部分是开发用于研究的新工具 氧化，生物系统中的一个基本过程。C)新功能 小的，高电子密度的电子显微镜标签。方法 将被开发用于在我们的新的 基于杂多钨酸盐(HPT)的EM标签，允许独立 生物系统中标记水平的量化。我们打算 合成光标、HPT标记的三磷酸腺苷 旨在确定三磷酸腺苷结合位点位置的研究 在F0F1-ATPase中，一种普遍存在的ATP合成酶。D)综合 以及二硫代N-丙氧基氮氧化物自旋标记的应用。 目的是提高化学稳定性的敏感度 我们实验室开发的丙氧基氮氧化物自旋标记，我们打算 目的：开发一条简便的合成全氢-15N-丙氧基的合成路线。 化学结构明确的自旋标记磷脂酰肌醇(PI) 以及一种标记的POSPONE类似物将被合成用于 脂质-蛋白的协同生物物理研究 PI与膜相关蛋白之间的相互作用 通过膜受体转导。