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）：通过天然存在的机制（例如磷酸化）对酶的调节是生命的基础。酶的小分子控制是许多药物作用的基础。酶活性的整体热控制可实现许多实验室和工业过程，例如 PCR 和阿斯巴甜合成。我们的总体目标是建立一种全新的酶调节方法。该方法利用金属纳米团簇（例如Au102(SR)44）在射频辐射下产生的纳米局部热量来热影响纳米团簇/酶缀合物的活性。选择 RF 时应尽量减少与混合物中其他成分的相互作用，类似于用于 Wi-Fi 的 RF。因此，这种纳米局部热酶控制不会改变溶液温度，也不应影响未直接缀合至纳米簇的酶的活性。我们建议通过 4 个具体目标来实现这一目标。目标 1 测试了我们可以在酶具有最小可测量活性的测定温度下“激活”嗜热酶/纳米簇缀合物（例如嗜热菌蛋白酶/Au102(SR)44）的假设。目标 2 检验了我们可以通过可逆解折叠来可逆地“失活”酶的假设。对于这一假设，我们首先测试纳米簇与溶菌酶、RNAse A 和 B-半乳糖苷酶等教科书酶的缀合物。在目标 1 和 2 中，我们测试了酶上纳米簇缀合位点影响缀合物活性的子假设。这种远程控制酶学的全面实施需要定量了解纳米团簇如何在射频下加热。这种理解将允许准确预测纳米簇温度，这是了解酶局部热度的先决条件。目前，提出了三种纳米团簇加热机制。它们是感应机制、磁机制和电泳机制。所有机制对所应用的射频场的频率都有不同的响应。目标 3 是测量少量明确的纳米团簇和纳米颗粒的频率响应。目标 4 是综合改变纳米粒子的特性，从而改变纳米粒子在不同机制下的热耗散。目标 3 和 4 都结合了使用现有机制的理论建模，认识到可能需要组合机制或开发一种新的理论机制来理解热耗散。这种机制的理解不仅将实现远程酶控制的新领域，而且还可能实现其他基于纳米颗粒的高温方法，例如非侵入性高温癌症治疗和远程控制分子生物物理学。