Catalogue of Human Salivary Secretory Components

人类唾液分泌成分目录

• 批准号: 6950761
• 负责人: John R Yates III
• 金额: $ 118.35万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-09-21 至 2008-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6950761
• 关键词: antibody; bioinformatics; clinical research; combinatorial chemistry; cooperative study; electrospray ionization mass spectrometry; human subject; hydroxyapatites; immunoaffinity chromatography; immunologic substance development /preparation; intermolecular interaction; liquid chromatography mass spectrometry; mass spectrometry; matrix assisted laser desorption ionization; oral health; peptide library; posttranslational modifications; protein purification; protein quantitation /detection; protein structure; protein structure function; proteomics; saliva; small molecule

DESCRIPTION (provided by applicant): Saliva is a complex mixture of proteins, lipids and carbohydrates in a physiological buffer containing millimolar concentrations of inorganic phosphate, divalent cations (calcium) and bicarbonate, which supports intermolecular complex formation, tooth mineralization, dental plaque formation, and passive immunity to pathogens of the oral cavity. We propose a four-year proteomics-based research program with the goal to identify and catalogue the components of salivary secretions and to annotate the catalogue with respect to the biochemical properties of the catalogue's inventory. To accomplish high coverage of the proteome, we will implement two parallel approaches to purify and identify individual salivary proteins. First, conventional protein fractionation will be coupled with either multi-dimensional liquid chromatography-tandem electrospray mass spectrometry (LC-MS/MS) or with 2-dimensional geI-MALDI-TOF mass spectrometry. Because the abundant salivary proteins will obscure the purification and identification of medium and low copy salivary proteins, a second purification technology, based on high-throughput immuno-affinity chromatography, will be implemented to complement and support the conventional mass spectrometry-driven proteomics approach. Specifically, we propose to create a comprehensive library of anti-saliva antibodies, using bacteriophage display, to build multiple probes for each salivary component. These antibody or immune affinity reagents will permit detection, as well as, purification of specific salivary proteins, which will, in turn, permit a high-throughput approach to the characterization of each protein's post-translational modification state, degree of interaction with other salivary components, and affinity for an in vivo tooth model (hydroxyapatite).

描述（由申请人提供）：唾液是蛋白质，脂质和碳水化合物的复杂混合物，其中包含毫米浓度的无机磷酸盐，二价阳离子（钙）和二氯苯甲酸盐，可支持分子矿物质矿化，牙齿矿化，牙齿矿物质，牙齿式不受欢迎，且普遍存在的无剂量对剂量的不受欢迎。 我们提出了一项为期四年的基于蛋白质组学的研究计划，其目标是识别和分类唾液分泌物的组成部分，并就目录库存的生化特性注释目录。 为了完成蛋白质组的高覆盖范围，我们将采用两种平行方法来净化和识别单个唾液蛋白。 首先，常规的蛋白质分馏将与多维液相色谱串联电喷雾质谱法（LC-MS/MS）或2维地质量 - 马尔迪-TOF质谱法结合。 由于丰富的唾液蛋白会掩盖中和低拷贝唾液蛋白的纯化和鉴定，因此将基于高通量免疫亲和力色谱法实施第二种纯化技术，以补充和支持常规的质谱群驱动的蛋白质组学方法。 具体来说，我们建议创建一个全面的 使用噬菌体显示的抗核抗体库，为每种探针构建多个探针 唾液成分。 这些抗体或免疫亲和力试剂将允许检测以及 纯化特定的唾液蛋白，这将允许对 每种蛋白质的翻译后修饰状态的表征，与 其他唾液成分以及对体内牙齿模型（羟基磷灰石）的亲和力。