Transient Induced Molecular Electronic Spectroscopy (TIMES) for study of protein-ligand interactions

用于研究蛋白质-配体相互作用的瞬态诱导分子电子光谱 (TIMES)

• 批准号: 1610516
• 负责人: yuhwa lo
• 金额: $ 39.52万
• 依托单位: University of California-San Diego
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-06-01 至 2020-03-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1610516&HistoricalAwards=false
• 关键词: Transient; Induced; Molecular; Electronic; Spectroscopy

Title:Transient Induced Molecular Electronic Spectroscopy (TIMES) for study of protein-ligand interactionsProject Goal:To test protein-ligand interactions, the key mechanisms for most drugs, without any external disturbances caused by molecular labelling or immobilization.Nontechnical Abstract:Protein ligand interaction is one of the most fundamental and widely studied areas in biology and chemistry because of their close relation to many diseases and disease therapies. Nearly all diseases including cancer, neural disorders, infectious diseases, cardiovascular diseases, renal diseases, metabolic diseases, immune diseases, etc. are connected to the abnormality of protein-protein or protein-ligand interactions. Unfortunately, all the current methods for in-vitro protein-ligand tests use either labels (often fluorescent) or ligand immobilization, both of which can produce substantial disruptions and introduce artifacts. These limitations have caused significant delay and cost increase in drug development. A new method is proposed and investigated to fundamentally solve the above problems, allowing precise in-vitro tests of the binding strength and kinetics of proteins ligand interactions to accelerate the process of drug discovery. The work can have significant impact on public health and the country's healthcare by shortening the time and saving the cost for drug discovery. The proposed research will also contribute to education and training of new generation of scientists for multi-disciplinary research involving biology, biochemistry, biophysics and engineering. The research also includes significant outreach activities to engage middle and high school students and underrepresented minority students to advance the STEM efforts.Technical Abstract:Since a protein and ligand have a large molecular weight and size difference (e.g.,100 kDa versus 1 kDa), linking organic fluorescent molecules to a protein or ligand can perturb or even change the binding properties. Similarly, the bioactivity of protein depends on its 3D configuration. Immobilization of the molecules limits its degree of freedom for binding, thus often yielding results different from reactions in physiological conditions. Lacking a precise method of in-vitro detection of protein-ligand interactions without any external disturbances has presented a major challenge for mid to late stage drug tests. Drug companies have often conducted unnecessary and unsuccessful human trials while the failures should have been detected earlier if a precise in-vitro test method exists. The most significant scientific and technological contributions of the proposed research will be the development of the method of Transient Induced Electronic Molecular Spectroscopy (TIMES) that allows label-free and immobilization-free detection of protein-ligand interactions. The TIMES method measures the signal caused by the dipole moment change when protein and ligand form protein-ligand complex. When integrated with ultralow noise electronics and microfluidics, the new µTIMES system will enable researchers to collect unprecedented rich information with high timing resolution and enhanced signal-to-noise ratio. This method will be applied to quantitative studies of protein-ligand and protein-aptamer interactions by measuring the dissociation constant and reaction kinetics.

标题：瞬态诱导分子电子光谱（TIMES）用于蛋白质-配体相互作用的研究项目目标：在没有分子标记或固定化引起的任何外部干扰的情况下，测试蛋白质-配体相互作用，这是大多数药物的关键机制。几乎所有的疾病，包括癌症、神经障碍、传染病、心血管疾病、肾脏疾病、代谢疾病、免疫疾病等，都与蛋白质-蛋白质或蛋白质-配体相互作用的异常有关。不幸的是，所有目前的体外蛋白质-配体测试方法都使用标记（通常是荧光）或配体固定，这两种方法都可能产生实质性的破坏并引入伪影。 这些限制导致药物开发的显著延迟和成本增加。 提出并研究了一种新的方法，以从根本上解决上述问题，允许精确的体外测试蛋白质配体相互作用的结合强度和动力学，以加速药物发现的过程。 这项工作可以通过缩短药物发现的时间和节省药物发现的成本，对公共卫生和国家的医疗保健产生重大影响。建议的研究亦有助教育和培训新一代科学家，从事涉及生物、生物化学、生物物理和工程的多学科研究。 该研究还包括重要的外展活动，以吸引初中和高中学生和代表性不足的少数民族学生，以推进STEM的努力。技术摘要：由于蛋白质和配体具有很大的分子量和大小差异（例如，100 kDa对1 kDa），将有机荧光分子连接到蛋白质或配体可以干扰甚至改变结合特性。同样，蛋白质的生物活性取决于其3D构型。分子的固定化限制了其结合的自由度，因此通常产生与生理条件下的反应不同的结果。缺乏一种精确的体外检测蛋白质-配体相互作用的方法，而没有任何外部干扰，这对中期到后期的药物测试提出了一个重大挑战。制药公司经常进行不必要的和不成功的人体试验，而如果存在精确的体外测试方法，失败应该更早被发现。拟议研究的最重要的科学和技术贡献将是瞬态诱导电子分子光谱（TIMES）方法的发展，该方法允许无标记和无固定化检测蛋白质-配体相互作用。TIMES方法测量当蛋白质和配体形成蛋白质-配体复合物时由偶极矩变化引起的信号。当与超低噪声电子器件和微流体集成时，新的µTIMES系统将使研究人员能够以高时序分辨率和增强的信噪比收集前所未有的丰富信息。该方法可用于蛋白质-配体和蛋白质-适体相互作用的定量研究。