Recognition and Presentation of Alpha Helices using Nanoparticle Receptors

描述（由申请人提供）：蛋白质的表面识别提供了新的治疗和诊断策略。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靶向螺旋裂缝蛋白质-蛋白质相互作用，如p53-HDM 2。在这些受体中，纳米颗粒单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）和将其模板化到靶分子的能力。在我们的研究中，我们将采用两种不同的策略：1）呈现螺旋以靶向互补蛋白质裂缝（例如HDM 2）; 2）将纳米颗粒模板化为螺旋以识别暴露的螺旋。本研究有三个具体目的。这些目标是相互关联的，需要综合、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模板？-螺旋，重点是螺旋识别的优化。目标二：共价键连接在目标1中开发的超分子肽-颗粒组装体，并将这些缀合物靶向含裂缝的蛋白质，包括HDM 2的p-53结合结构域。这些研究将侧重于：1）颗粒稳定肽的螺旋结构的能力; 2）使用颗粒单层来增强受体的亲和力并控制结合蛋白的结构。目标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肽。肽与颗粒的复合将淬灭荧光团。然后，在肽被靶蛋白置换后的荧光增强将用于抑制结合。蛋白质-颗粒结合过程的选择性将以阵列形式应用，以提供蛋白质的“化学鼻”感测。相关性：这项研究的重点是创造潜在的抗癌疗法和检测用于诊断癌症和其他疾病状态的蛋白质生物标志物。

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.