Radiofrequency Remote Control of Enzyme-Nanocluster Conjugates

酶-纳米团簇缀合物的射频远程控制

• 批准号: 9061746
• 负责人: Christopher Jeffries Ackerson
• 金额: $ 27.8万
• 依托单位: COLORADO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-05-01 至 2019-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9061746
• 关键词: Accounting; Address; Amyloid; Aspartame; Bacillus (bacterium); Biological; Biological Assay; Biological Process; Biophysics; Budgets; Caliber; Charge; Coupled; Dependence; Dependency; Development; Enzymatic Biochemistry; Enzyme Activation; Enzyme Kinetics; Enzyme Tests; Enzymes; Frequencies; Galactosidase; Generations; Goals; Health; Heating; Label; Laboratories; Life; Magnetism; Measurable; Measures; Metals; Methods; Modeling; Molecular; Muramidase; Nature; Pathway interactions; Pharmaceutical Preparations; Pharmacologic Substance; Phosphoglycerate Kinase; Phosphorylation; Physical Chemistry; Process; Property; Proteins; Radial; Radiation; Regulation; Resolution; Running; Site; Structure; Surface; Temperature; Testing; Textbooks; Theoretical model; Thermolysin; Thermus thermophilus; Work; base; cancer therapy; density; electric field; enzyme activity; enzyme model; insight; irradiation; magnetic field; nano; nanomaterials; nanoparticle; nanoscale; novel; oxidation; particle; physical property; protein complex; radiofrequency; response; small molecule; standard measure; thermophilic organism

DESCRIPTION (provided by applicant): Regulation of enzymes by naturally occurring mechanisms such as phosphorylation is fundamental to life. Small molecule control of enzymes underlies the action of many pharmaceutical drugs. Bulk thermal control of enzymatic activity enables many laboratory and industrial processes such as PCR and aspartame synthesis. Our overall goal is to establish an entirely new method for regulating enzymes. This method uses the nano- localized heat generated by metal nanoclusters such as Au102(SR)44 under radiofrequency irradiation to thermally influence the activity of a nanocluster/enzyme conjugate. The RF is chosen to interact minimally with other components of the mixture, analogous to the RF used for Wi-Fi. Thus, this nano-local thermal enzyme control does not modify the solution temperature, nor should it influence the activity of enzymes that are not directly conjugated to nanoclusters. We propose to accomplish this goal in a set of 4 specific aims. Aim 1 tests the hypothesis that we can 'activate' thermophilic enzyme/nanocluster conjugates such as thermolysin/Au102(SR)44 at assay temperatures in which the enzyme has minimal measurable activity. Aim 2 tests the hypothesis that we can reversibly 'deactivate' enzymes, presumably by reversible unfolding. For this hypothesis we begin by testing nanocluster conjugates to textbook enzymes such as lysozyme, RNAse A and B-galactosidase. In both Aims 1 and 2 we test the sub-hypothesis that the site of nanocluster conjugation on the enzyme influences the activity of the conjugate. A full implementation of this remote-control enzymology requires quantitative understanding of how nanoclusters heat in radiofrequencies. Such an understanding will allow accurate prediction of nanocluster temperature, which is prerequisite for understanding how locally hot an enzyme is. Currently, there are three proposed mechanisms for nanocluster heating. These are an inductive mechanism, a magnetic mechanism, and an electrophoretic mechanism. All mechanisms have different responses to the frequency of the applied radiofrequency filed. Aim 3 is to measure the frequency response of a small number of well-defined nanoclusters and nanoparticles. Aim 4 is to synthetically change the properties of nanoparticles in a manner that will change their thermal dissipation under different mechanisms. Both aims 3 and 4 incorporate theoretical modeling using existing mechanisms, recognizing the possible need for combining mechanisms or developing a novel theoretical mechanism for understanding thermal dissipation. Such mechanistic understanding will not only enable a new field of remote enzyme control but may also enable other nanoparticle based hyperthermal methods such as noninvasive hyperthermal cancer therapy and a remote control molecular biophysics.

描述（由申请人提供）：通过自然发生的机制（例如磷酸化）对酶的调节是生命的基础。小分子对酶的控制是许多药物的作用。酶活性的批量热控制使许多实验室和工业过程（例如PCR和Aspartame合成）。我们的总体目标是建立一种调节酶的全新方法。该方法使用金属纳米簇产生的纳米局部热量，例如Au102（SR）44在辐射频率辐照下，从而热影响纳米簇/酶结合的活性。选择RF与混合物的其他成分相互作用，类似于用于Wi-Fi的RF。因此，这种纳米局部热酶对照不会改变溶液温度，也不应影响未直接偶联与纳米簇的酶的活性。我们建议在一组4个特定目标中实现这一目标。 AIM 1检验了一个假设，即我们可以在测定温度下“激活”嗜热酶/纳米簇共轭物，例如热溶胶蛋白/AU102（SR）44，其中该酶具有最小的可测量活性。 AIM 2检验了我们可以可逆地“失活”酶的假设，大概是通过可逆的展开来进行的。对于这一假设，我们首先测试纳米簇共轭物，以溶菌酶，RNase A和B-半乳糖苷酶等教科书酶。在两个目标1和2中，我们测试了纳米簇共轭的位点在酶上影响结合物的活性。这种遥控酶的充分实施需要定量了解纳米群体如何在放射性频率中加热。这种理解将允许对纳米簇温度进行准确的预测，这是理解酶在局部热的前提。当前，有三种提出的纳米簇加热的机制。这些是电感机制，一种磁性机制和电泳机制。所有机制对提交的辐射射频的频率都有不同的响应。 AIM 3是测量少数定义明确的纳米簇和纳米颗粒的频率响应。目的4是以一种将在不同机制下改变其热耗散的方式来合成更改纳米颗粒的性质。两者的目标3和4都使用现有机制融合了理论建模，认识到结合机制或开发一种新的理论机制来理解热耗散的必要性。这种机械理解不仅将实现远程酶控制的新领域，而且还可以实现其他基于纳米颗粒的高温方法，例如非侵投性高温癌症治疗和遥控分子生物物理学。