Pathogen Recognition by Toll-like Receptors

Toll 样受体对病原体的识别

• 批准号: 6445092
• 负责人: JEFFREY A LAWTON
• 金额: $ 4.62万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-02-01 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6445092
• 关键词: CD14 molecule; CHO cells; Escherichia coli; Sf9 cell line; X ray crystallography; analytical ultracentrifugation; biological signal transduction; biophysics; communicable diseases; crystallization; fluorescent dye /probe; host organism interaction; intermolecular interaction; microorganism immunology; model design /development; molecular site; physical model; protein purification; protein structure function; structural biology; surface antigens; surface plasmon resonance; toll like receptor

DESCRIPTION (provided by applicant): Many cell types in vertebrates express proteins which are capable of detecting the presence of a diverse array of pathogens, including bacteria, mycoplasmas, and fungi, a capacity which is known as innate immunity. Upon recognition of invading pathogens, these cells produce specific inflammatory cytokines which signal the immune system to mount a response. A small class of proteins called toll-like receptors (TLRs) has recently been shown to play a central role in transducing the signal from invading pathogens across the plasma membrane. The mechanism of pathogen recognition is not presently understood from a structural perspective. The objective of the research propsed here is to characterize the interaction between TLRs and various ligands using biophysical and structural approaches according to the following specific aims: (1) overexpression and purification of the extracellular domains of human TLR2 or TLR4 along with the adaptor proteins CD14 and MD-2; (2) biophysical characterization of the binding affinities of various ligands and adaptor proteins using analytical ultracentrifugation, fluorescence anisotropy, and surface plasmon resonance; (3) determination of the atomic structure of the extracellular domains of TLR2 or TLR4 using x-ray crystallography.

描述（由申请人提供）：脊椎动物中的许多单元格式 能够检测存在多种阵列的蛋白质 病原体，包括细菌，支原体和真菌，一种能力 被称为先天免疫。认识到入侵病原体后，这些细胞 产生特定的炎症细胞因子，这些细胞因子向免疫系统发出信号 回应。一小类称为Toll样受体（TLR）的蛋白质具有 最近被证明在传输信号中起着核心作用 跨质膜入侵病原体。病原体的机制 从结构的角度来看，目前尚未了解识别。这 这里支撑的研究的目的是表征相互作用 在TLR和各种配体之间使用生物物理和结构方法 根据以下特定目的：（1）过表达和纯化 人类TLR2或TLR4的细胞外结构域以及适配器 蛋白质CD14和MD-2; （2）结合的生物物理表征 使用分析的各种配体和衔接蛋白的亲和力 超速离心，荧光各向异性和表面等离子体共振； （3）测定TLR2细胞外域的原子结构 或使用X射线晶体学的TLR4。