Nanoscale Investigation of Protein-Surface Interactions

描述（由申请人提供）：肽和蛋白质在界面处的相互作用和分子结构在广泛的生物医学应用（植入生物材料、诊断阵列、细胞培养物、组织工程构建体等）中发挥重要作用。在这里，我们建议开发一套互补的表面生物分析和计算工具，以提供有关肽和蛋白质与表面的结构和相互作用的纳米级信息。最初，将使用具有明确定义的a-螺旋和b-折叠二级结构的模型亮氨酸/赖氨酸（LK）肽来开发该工具集。研究的表面范围从烷乙氧基化的纳米颗粒到纳米结构的基材。在这些研究中使用的实验技术将包括固态NMR、静态飞行时间二次离子质谱、和频产生、近边X射线吸收精细结构、表面等离子体共振和X射线光电子能谱。为了实现这些实验技术的全部功能，将开发互补的计算方法，使用分子动力学模拟从理论上分析这些相同系统的吸附行为。这些实验和计算方法将提供一个全面的了解，在纳米级的二级结构，分子取向和侧链/表面结合肽的表面相互作用，以及界面水在肽/表面相互作用和结构的作用。然后，这些方法将扩展到更复杂的生物分子，蛋白质G的纳米级表征。这个新工具集提供的信息将阐明在纳米级的明确结构中将肽和蛋白质附着在表面上的设计原理，从而实现蛋白质吸附和固定的分子水平控制。这将在生物应用中具有广泛的影响，因为在纳米级控制表面固定的肽和蛋白质的构象、取向等将直接影响生物医学装置的生物活性和生物相容性。将蛋白质和肽附着在纳米颗粒表面是早期检测和治疗癌症的一种有前途的策略。类似地，将蛋白质和肽附着到生物医学设备上有望改善这些设备的功能。本提案中开发的工具将提供最佳地将蛋白质和肽附着到表面所需的理解和信息。

DESCRIPTION (provided by applicant): The interactions and molecular structures of peptides and proteins at interfaces play an important role in a wide range of biomedical applications (implanted biomaterials, diagnostic arrays, cell cultures, tissue engineering constructs, etc.). Here we propose the development of a complementary suite of surface bioanalytical and computational tools to provide information at the nanoscale about the structures and interactions of peptides and proteins with surfaces. Initially model leucine/lysine (LK) peptides with well- defined a-helix and b-sheet secondary structures will be used to develop this tool set. The surfaces investigated will range from alkanethiol functionalized nanoparticles to nanostructured substrates. The experimental techniques used in these studies will include solid state NMR, static time-of-flight secondary ion mass spectrometry, sum frequency generation, near edge x-ray absorption fine structure, surface plasmon resonance and x-ray photoelectrdn spectroscopy. To realize the full power of these experimental techniques, complementary computational approaches will be developed to theoretically analyze the adsorption behavior of these same systems using molecular dynamics simulations. Together these experimental and computational methods will provide a comprehensive understanding at the nanoscale of the secondary structure, molecular orientation and side chain/surface interactions of surface bound peptides, and the role of interfacial water in the peptide/surface interactions and structures. Then, these methods will be extended to the nanoscale characterization of a more complex biomolecule, Protein G. The information provided by this new tool set will elucidate the design principles for attaching peptides and proteins on surfaces in well-defined structures at the nanoscale, enabling molecular-level control of protein adsorption and immobilization. This will have wide spread impact in biological applications since controlling the conformation, orientation, etc. of surface immobilized peptides and proteins at the nanoscale will directly effect bioactivity and biocompatibility of biomedical devices. The attachment of proteins and peptides to the surface of nanoparticles is a promising strategy for the early detection and treatment of cancer. Similarly, the attachment of proteins and peptides to biomedical devices holds the promise for improving the function of those devices. The tools developed in this proposal will provide understanding and information needed to optimally attach protein and peptides to surfaces.