Volume profile of biochemically responsive interfaces

生化响应界面的体积分布

• 批准号: 243193367
• 负责人: Professor Dr. Claus Czeslik
• 金额: --
• 依托单位: Forschungsgebiet Physikalische Chemie
• 依托单位国家: 德国
• 项目类别: Research Units
• 财政年份: 2013
• 资助国家: 德国
• 起止时间: 2012-12-31 至 2019-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/243193367?language=en
• 关键词: Volume; profile; biochemically; responsive; interfaces

Some proteins undergo major conformational changes upon ligand binding, such as calmodulin. This functionality shall be transferred to interfacial structures in this proposed project. Aqueous-solid interfaces are modified by multilayers or brushes of polyelectrolytes incorporating calmodulin or by covalent bonding of calmodulin mutants to the solid phase. In this way, novel biochemically responsive interfacial structures are formed, in contrast to well-known pH or temperature sensitive interfacial architectures. Such interfaces are relevant for many applications, where, for example, the immobilization of enzymes, the cell growth or the release of drugs needs to be regulated. Main goals of the proposed project are the controlled build-up of such interfacial layers and the characterization of the calmodulin functionality in these layers. Pressure is used as key variable to determine the volume changes associated with the build-up of the interfacial layers, the adsorption of protein and the ligand binding to calmodulin. Furthermore, the functionality of the biochemically responsive interfaces is investigated as a function of pressure, because pressure allows for tuning intermolecular interactions between protein and polyelectrolyte and modulating the packing density of the interfacial layers. The achievements and the methods used in the first funding period, where pressure effects on enzyme reactions in similar systems have been investigated, will be a helpful basis for the new proposed experiments. High-pressure surface-sensitive analytical tools will be applied, and similar interfacial modifications are prepared, complemented by covalent binding of calmodulin mutants. Moreover, surface modifications, which are biocompatible for enzyme reactions, have been identified in the first round of experiments. The results of this new project will describe the build-up, the thermodynamic and dynamic properties as well as the biochemical functionality of calmodulin-modified interfacial structures in detail, which is important for the understanding of these systems and their biotechnological and biomedical applications.

一些蛋白质在配体结合后经历主要构象变化，如钙调蛋白。该功能应转移到本拟议项目中的界面结构。水-固界面通过多层或刷状的聚电解质结合钙调蛋白或通过钙调蛋白突变体与固相的共价键合来修饰。以这种方式，形成了新的生物化学响应界面结构，与众所周知的pH或温度敏感的界面结构相反。这样的界面对于许多应用是相关的，其中，例如，酶的固定化、细胞生长或药物的释放需要被调节。拟议项目的主要目标是控制建立这样的界面层和这些层中的钙调素功能的表征。压力被用作关键变量，以确定与界面层的建立，蛋白质的吸附和配体结合到钙调蛋白的体积变化。此外，作为压力的函数的生化响应接口的功能性进行了研究，因为压力允许调整蛋白质和蛋白质之间的分子间相互作用和调制的界面层的堆积密度。在第一个资助期内，研究了类似系统中压力对酶反应的影响，所取得的成就和使用的方法将为新提议的实验提供有益的基础。将应用高压表面敏感的分析工具，并准备类似的界面修饰，辅之以钙调素突变体的共价结合。此外，在第一轮实验中已经确定了对酶反应具有生物相容性的表面修饰。这个新项目的结果将详细描述钙调素修饰的界面结构的建立，热力学和动力学性质以及生化功能，这对于理解这些系统及其生物技术和生物医学应用非常重要。