Finding the right niche:quantifying protein folding stability in materials

这一计划提出了一种创新的方法，直接量化材料属性的影响 在亚微米空间分辨率和毫秒时间分辨率下的原位蛋白质折叠稳定性。一个 设计含蛋白质组分的杂化生物材料的主要挑战是 造成蛋白质活性的大量损失，具有重大的商业、科学和临床意义 后果。解决这一问题的一个主要障碍是无法对材料进行现场量化 保留或关闭蛋白质功能的特性。本程序将开发一种动态荧光 成像方法，快速松弛成像(FREI)，有望克服这一障碍并打开 这是在蛋白质水平上对蛋白质-物质相互作用进行新研究的大门。此程序构建 根据我们最近的演示，FREI测量能够量化蛋白质折叠的稳定性 聚丙烯酰胺水凝胶。我们现在试图将这种方法扩展到与生物医学相关的研究 蛋白质和材料。具体目标1将通过以下方式测试识别物理化学机制的能力 哪些材料扰乱了蛋白质的稳定性和功能，通过使用二维底物来控制 表面化学。特殊目标2将把研究扩展到三维包裹的蛋白质 藻酸盐水凝胶，广泛用于药物输送和组织工程。结果来自 该计划和预期的后续研究将为产生新的分子奠定基础 具有生物大分子成分的坚固、功能生物材料的水平设计规则。

This program advances an innovative approach that directly quantifies the impact of material properties on protein folding stability in situ at sub-micron spatial resolution and at millisecond time resolution. A major challenge in engineering hybrid biomaterials with protein components is that many materials contribute to substantial losses of protein activity, with significant commercial, scientific, and clinical ramifications. A major roadblock to solving this problem is the inability to quantify in situ the material properties that preserve or shut down protein function. This program will develop a dynamic fluorescence imaging approach, Fast Relaxation Imaging (FReI) that promises to overcome this roadblock and open the door to novel investigations of protein-material interactions, at the protein level. This program builds on our recent demonstration that FReI measurements are able to quantify protein folding stability in polyacrylamide hydrogels. We now seek to extend this approach to studies of biomedically relevant proteins and materials. Specific Aim 1 will test the capacity to identify physical chemical mechanisms by which materials perturb protein stability and function, by using two-dimensional substrates of controlled surface chemistry. Specific Aim 2 will extend studies to encapsulated proteins in three-dimensional alginate hydrogels, which are extensively used for drug delivery and tissue engineering. Outcomes from this program and anticipated subsequent research will lay the foundation for generating new molecular level design rules for robust, functional biomaterials with bio-macromolecular components.