Identifying Key Structural Interactions in Heparan Sulfate-Protein Complexes

鉴定硫酸乙酰肝素-蛋白质复合物中的关键结构相互作用

• 批准号: 10715985
• 负责人: Kari Joanne Pederson
• 金额: $ 16.27万
• 依托单位: CALIFORNIA STATE UNIV-DOMINGUEZ HILLS
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-01 至 2027-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10715985
• 关键词: Autoimmune Diseases; Binding; Binding Proteins; Biological; Biological Process; Blood Circulation; Blood Coagulation Factor; CCL2 gene; CXCL10 gene; California; Calorimetry; Carbohydrates; Catalysis; Categories; Cell Adhesion; Cell Adhesion Molecules; Cell Communication; Cell Culture Techniques; Cell surface; Cells; Characteristics; Coagulation Process; Collaborations; Complex; Core Protein; Data; Disaccharides; Disease Progression; Docking; Drug Targeting; Early identification; Extracellular Matrix; Foundations; Funding; Future; Glycosaminoglycans; Goals; Growth Factor; HIV-1; Heparitin Sulfate; Hispanic-serving Institution; Human; IL8 gene; Immune response; Infection; Infection prevention; Inflammation; Isotope Labeling; Libraries; Life Cycle Stages; Ligands; Mammalian Cell; Membrane Proteins; Methodology; Modeling; Molecular; Molecular Conformation; Nuclear Magnetic Resonance; Process; Proteins; Proteoglycan; RANTES; Research; Role; Signal Transduction; Spectrum Analysis; Stromal Cell-Derived Factor 1; Structural Models; Structural Protein; Structure; Surface Plasmon Resonance; Testing; Titrations; Training; Universities; Virus; Virus Diseases; cell growth regulation; chemokine; cytokine; drug development; experimental analysis; extracellular; institutional capacity; nuclear transfer; programs; protein complex; receptor; screening; sugar; tissue repair; undergraduate student; viral entry inhibitor; virtual

Project Summary Glycosaminoglycans (GAGs), such as heparan sulfate (HS), are important components of the extracellular matrix of all cells. The extraordinary structural diversity of GAGs enables them to interact with a wide variety of biological molecules, but primarily proteins, to modulate biological processes, such as cell adhesion, tissue repair, coagulation, and immune response, among many others. Additionally, a number of different viruses, including HIV-1, initiate infection of cells by binding to cell surface GAGs. Unfortunately, the diversity of potential binding motifs in GAG sequences has made identification of relevant sequences challenging. In particular, heparan sulfate (HS) is the most structurally diverse, with sixteen unique disaccharide configurations and 256 unique tetrasaccharide configurations. Significant progress has been made in recent years developing libraries of synthetic HS oligomers, allowing for detailed experimental structural studies of HS-protein complexes. Understanding the structural basis of these interactions is key to drug development and the HS binding protein-HS (HSBP-HS) interaction studies to date have been either non-structural or computational. The overall goal of this project is to utilize these recently developed synthetic HS libraries and NMR-based structural models to identify key interactions. HIV-1 p17 will be used for initial studies because it is very important in many stages of the life cycle of HIV-1. Heparan sulfate receptors are known to influence growth factor signaling, cell adhesion, and enzymatic catalysis on human cells. Thus, this interaction may be one of the deciding factors for disease progression in humans infected with HIV-1. In Aim 1, a library of synthetic HS tetrasaccharides will be screened for binding to human immunodeficiency virus type 1 (HIV-1) p17 protein to identify the sequence-specific HS characteristics of the preferred binding oligomers. In Aim 2, the HSBP-HS complexes of both strong and weak binding sequences will be studied via nuclear magnetic resonance (NMR) to model the structure of the complex and identify key interactions for future drug development. After the methodology is established, HS-binding chemokines will be screened in Aim 3 to identify suitable targets for future NMR studies.

项目概要 糖胺聚糖 (GAG)，例如硫酸乙酰肝素 (HS)，是细胞外基质的重要组成部分。 所有细胞的矩阵。 GAG 非凡的结构多样性使它们能够与多种相互作用 生物分子，但主要是蛋白质，调节生物过程，例如细胞粘附、组织 修复、凝血和免疫反应等。此外，许多不同的病毒， 包括 HIV-1，通过与细胞表面 GAG 结合来启动细胞感染。不幸的是，多样性 GAG 序列中潜在的结合基序使得相关序列的识别具有挑战性。在 特别是，硫酸乙酰肝素 (HS) 结构最多样化，具有 16 种独特的二糖构型 和 256 种独特的四糖构型。近年来发展取得显着进展 合成 HS 寡聚物文库，可对 HS 蛋白进行详细的实验结构研究 复合物。了解这些相互作用的结构基础是药物开发和 HS 的关键 迄今为止，结合蛋白-HS (HSBP-HS) 相互作用研究要么是非结构性的，要么是计算性的。 该项目的总体目标是利用这些最近开发的合成 HS 库和基于 NMR 的 识别关键相互作用的结构模型。 HIV-1 p17 将用于初步研究，因为它非常有效 在 HIV-1 生命周期的许多阶段都很重要。已知硫酸乙酰肝素受体会影响生长 人体细胞的因子信号传导、细胞粘附和酶催化。因此，这种相互作用可能是以下之一： 感染 HIV-1 的人类疾病进展的决定因素。在目标 1 中，合成 HS 库 将筛选与人类免疫缺陷病毒 1 型 (HIV-1) p17 蛋白结合的四糖，以 确定首选结合寡聚体的序列特异性 HS 特征。在目标 2 中，HSBP-HS 将通过核磁共振（NMR）研究强结合序列和弱结合序列的复合物 对复合物的结构进行建模并确定未来药物开发的关键相互作用。之后 方法建立后，将在目标 3 中筛选 HS 结合趋化因子，以确定合适的靶标 未来的核磁共振研究。