Recognition and Presentation of Alpha Helices using Nanoparticle Receptors

描述(由申请人提供)：蛋白质的表面识别为新的治疗和诊断策略提供了途径。在我们提出的研究中，我们将使用基于金纳米颗粒的合成受体来针对螺旋-裂解蛋白-蛋白质相互作用，例如P53-Hdm2。在这些受体中，纳米颗粒单分子层将在识别过程中发挥积极作用，利用纳米颗粒的尺寸尺度(直径为6-10 nm)和纳米颗粒作为目标分子的模板的能力。在我们的研究中，我们将采取两种不同的策略：1)提供针对互补蛋白裂隙的螺旋(例如Hdm2)；2)将纳米颗粒模板化为螺旋以识别暴露的螺旋。这项拟议的研究有三个具体目标。这些目标是相互关联的，运用了合成、物理和生物物理方法。在这些目标中，我们将：目标1：确定纳米粒子静电结合的范围和事件及其与β-螺旋的模板，重点是螺旋识别的优化。目的2：共价连接目标1中开发的超分子多肽-颗粒组合体，并将这些连接物靶向裂隙蛋白，包括hdm2的p-53结合域。这些研究将集中在：1)颗粒稳定多肽螺旋结构的能力；2)利用颗粒单层来增强受体的亲和力和控制结合蛋白的结构。目的3：使用纳米颗粒作为生物医学相关蛋白质的传感器。在这些研究中，颗粒将被模板化为荧光团标记的多肽。多肽与微粒的络合作用将使荧光团熄灭。然后，目标蛋白取代多肽时的荧光增强将用于转导结合。蛋白质-颗粒结合过程的选择性将以阵列形式应用，以提供对蛋白质的“化学鼻子”传感。相关性：这项研究的重点是创造潜在的抗癌疗法，并检测用于诊断癌症和其他疾病状态的蛋白质生物标记物。

DESCRIPTION (provided by applicant): Surface recognition of proteins provides access to new therapeutic and diagnostic strategies. In our proposed research we will target helix-cleft protein-protein interactions such as p53-HDM2 using gold nanoparticle-based synthetic receptors. In these receptors, the nanoparticle monolayer will play an active role in the recognition process, exploiting the size scale (6-10 nm in diameter) and the ability of nanoparticles to be templated to target molecules. We will pursue two different strategies in our research: 1) Presentation of helices for the targeting of complementary protein clefts (e.g. HDM2); 2) Templation of the nanoparticles to helices for recognition of exposed helices. The proposed research features three specific Aims. These aims are interrelated, bringing to bear synthetic, physical and biophysical methodologies. In these Aims, we will: Aim 1: Determine the scope and events involved in electrostatic binding of nanoparticles and their templation to ?-helices, focusing on the optimization of helix recognition. Aim 2: Covalently link the supramolecular peptide-particle assemblies developed in Aim 1, and target these conjugates to cleft-bearing proteins, including the p-53 binding domain of HDM2. These studies will focus on: 1) The ability of the particle to stabilize the helical structure of the peptide; 2) The use of the particle monolayer to enhance the affinity of the receptors and to control the structure of the bound protein. Aim 3: Use nanoparticles as sensors for biomedically-relevant proteins. In these studies, particles will be templated to fluorophore-tagged peptides. The complexation of the peptide by the particle will quench the fluorophore. Fluorescent enhancement upon displacement of the peptide by the target protein will then be used to transduce binding. The selectivity of the protein-particle binding process will be applied in an array format to provide "chemical nose" sensing of proteins. Relevance: This research focuses on the creation of potential anti-cancer therapeutics and the detection of protein biomarkers for diagnosis of cancer and other disease states.