UNDERSTANDING THE INTERACTION OF ANKYRINS AND NANOMATERIALS

描述（由申请人提供）：蛋白质与固体表面的相互作用是一种基本现象，对纳米技术、生物材料和生物技术工艺具有影响。虽然“大多数蛋白质与大多数表面相互作用”，但每种相互作用的特定强度、机制和动力学对吸附蛋白质的最终构象具有重要影响。吸附的基础通常由疏水和静电相互作用的某种组合提供。虽然通常被认为是驱动结合的有利自由能变化的主要贡献者，但疏水相互作用诱导吸附蛋白质中生物活性的显著重排和损失。静电相互作用虽然通常太弱而不能提供长期稳定性，但能够保持蛋白质结构，并且可以通过多种方式进行控制。因此，我们建议1）利用静电力来控制蛋白质吸附到电极表面的过程，2）通过调节施加到表面的电势来操纵吸附的蛋白质的活性。该项目的假设是，通过控制施加的电势， 将蛋白质吸附到表面（电极）上，可能会影响吸附过程（亲和性、机理和动力学），最重要的是，影响吸附蛋白质的生物活性。目前的证据，虽然只有很少的报道，主要是定性表达，支持这一假设。因此，本项目的主要目标是系统地证明（并了解如何，为什么，以及多快）电极电位的变化可以影响吸附，取向，构象，活性和吸附蛋白质的稳定性。 对于这些研究，我们选择了一组称为锚蛋白的蛋白质。这些蛋白质显示出异常高的稳定性和完全可逆的弹簧样行为。除了目前还没有这类蛋白质吸附行为的相关报道外，本研究还将确定表面电位在吸附和最终构象中的基本机制，这对于支持传感器和纳米器件的合理开发和应用至关重要。此外，了解锚蛋白和其他膜蛋白如何与纳米材料相互作用将成为开发高通量模型的基础，以研究与缺陷蛋白质-蛋白质相互作用相关的广泛病理学，如遗传性球形红细胞增多症，脊髓小脑共济失调，心律失常和各种通道病。拟议的项目还将开发一种新的基于表面的方法，以研究在药物化合物存在下蛋白质的实时结合，否则这些药物化合物会干扰细胞中其他蛋白质的结合。

DESCRIPTION (provided by applicant): The interaction of proteins with solid surfaces is a fundamental phenomenon with implications on nanotechnology, biomaterials and biotechnological processes. Although "most proteins interact with most surfaces", the particular strength, mechanism, and kinetics of each interaction has significant consequences in the final conformation of the adsorbed protein. The basis for the adsorption is generally provided by some combination of hydrophobic and electrostatic interactions. While generally recognized as the major contributor to the favorable free energy change driving the binding, hydrophobic interactions induce significant rearrangements and losses in biological activity in the adsorbed protein. Electrostatic interactions, though typically too weak to provide long-term stability, enabe preserving the protein structure and can be controlled by a variety of ways. Hence, we propose to 1) exploit electrostatic forces to control the adsorption process of proteins to electrode surfaces and 2) manipulate the activity of the adsorbed proteins by adjusting the potential applied to the surface. The hypothesis of this project is that by controlling the potential applied to the surface (electrode), it will be possible to affect the adsorption process (affinity, mechanism, and kinetics) and most importantly, the biological activity of the adsorbed proteins. Current evidence, though only sparsely reported and mostly qualitatively expressed, supports this hypothesis. Thus, the main goal of this project is to systematically demonstrate that (and understand how, why, and how fast) changes in electrode potential can affect the adsorption, orientation, conformation, activity, and stability of adsorbed proteins. For these studies, we have selected a group of proteins called ankyrins. These proteins display an unusually high stability and fully reversible spring-like behavior. Besides the fact that there are no reports related to adsorption behavior of these proteins, this proposal will determine the fundamental mechanism of surface potential in the adsorption and final conformation, which is crucial to bolster the rational development and application of sensors and nanodevices. Furthermore, understanding how ankyrins and other membrane proteins, interact with nanomaterials will form the basis for the development of a high-throughput model to study a broad range of pathologies linked to defective protein-protein interactions, such as hereditary spherocytosis, spinocerebellar ataxia, cardiac arrhythmias, and a variety of channelopathies. The proposed project will also develop a novel surface-based method to study real-time binding of proteins in the presence of pharmaceutical compounds that would otherwise interfere with the association of other proteins in cells.