Core B: Mucus/Mucin Analytics Core

核心 B：粘液/粘蛋白分析核心

• 批准号: 10684187
• 负责人: Mehmet Kesimer
• 金额: $ 26.83万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-15 至 2027-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10684187
• 关键词: Adhesions; Agar Gel Electrophoresis; Biochemical; Biophysics; Cell Culture Techniques; Charge; Cilia; Coupled; Disease; Doctor of Philosophy; Dryness; Electron Microscopy; Epithelium; Gel; Gland; Goblet Cells; Health; Human; Human Resources; Investigation; Label; Laboratories; Lasers; Left; Liquid substance; MUC5AC gene; MUC5B gene; Mass Spectrum Analysis; Measurement; Measures; Medicine; Modeling; Molecular; Molecular Sieve Chromatography; Molecular Weight; Mucin 1 protein; Mucin-2 Staining Method; Mucins; Mucous body substance; Mus; North Carolina; Pathology; Performance; Phenotype; Positioning Attribute; Property; Proteins; Proteome; Proteomics; Refractometry; Research Personnel; Resources; Respiratory Mucosa; Sampling; Services; Shapes; Solid; Specific qualifier value; Structure; Submucosa; Surface; Techniques; Training; Ultracentrifugation; Universities; Weight; Western Blotting; air sampling; dimer; in vivo; instrumentation; light scattering; molecular size; professor; respiratory; success; viscoelasticity; zeta potential

Abstract The Mucus/Mucin Analytics Core will perform key mucin/mucus biochemical, biophysical, and proteomics measurements for all the Projects of the PPG. These services are required for our PPG investigators who are focused on a mechanistic understanding of the physical and biochemical forces which govern mucus viscoelasticity, transport, and the adhesion of mucus to epithelial surfaces. Qualitative and quantitative characterization of mucus secretions from culture models, submucosal gland mucus, and Muc5b dimer and WT mice and their MTEC cultures are key for successful execution the projects and for the overall synergistic success of this PPG. Mucus and mucin measures raise unique challenges in their performance from biochemical, biophysical, and proteomic perspectives. Therefore, the mucus/mucin analytics core is a centralized resource for aiding all the PPG projects with their mucin and mucus analysis requirements. The Mucus/Mucin analytics Core will offer the following services to the Projects: 1- Mucin/Mucus quantitation: Total mucin concentrations, total biomolecule concentrations and solids (%) dry weight measurements of overall mucus concentration in mucus samples will be measured. Absolute concentration of mucins MUC5AC and MUC5B, and other airways innate defense proteins, including PRR4, LPLUNC1, FCGBP, will be measured by labelled mass spectrometry. 2- Mucin characterization: Isolation of mucin molecules and characterization of their macromolecular properties, including molecular weight (MW), hydrodynamic size (Rg), charge status, are key parameters for the necessary calculations made by the projects. Molecular weight and size, oligomerization status, shape (compact vs linear), charge status will be assessed by GPC, refractometry, multi-angle laser light scattering (MALLS), western blotting following agarose gel electrophoresis, and rate zonal ultracentrifugation followed by electron microscopy. 3- Mucin/Mucus proteome: Mass spectrometry-based proteomics characterization of culture secretions and, gland secretions, and the proteins comprising the mucin-interacting proteome will be identified.

抽象的 粘液/粘蛋白分析核心将执行关键的粘蛋白/粘液生化，生物物理和蛋白质组学 PPG所有项目的测量。这些服务是我们的PPG调查人员需要的 专注于对控制粘液的物理和生化力的机械理解 粘弹性，运输和粘液对上皮表面的粘附。定性和定量 培养模型，粘膜粘膜粘液以及MUC5B二聚体和WT的粘液分泌物表征 小鼠及其MTEC文化是成功执行项目和整体协同作用的关键 这个PPG的成功。粘液和粘蛋白措施在生化的表现中引起了独特的挑战， 生物物理和蛋白质组学的观点。因此，粘液/粘蛋白分析核心是一种集中资源 为了帮助所有PPG项目具有粘蛋白和粘液分析要求。粘液/粘蛋白分析 核心将为项目提供以下服务：1-粘蛋白/粘液定量：总粘蛋白浓度， 总生物分子浓度和固体（％）干重测量的总粘液浓度 将测量粘液样品。粘蛋白MUC5AC和MUC5B的绝对浓度以及其他气道 先天防御蛋白，包括PRR4，LPLUNC1，FCGBP，将通过标记的质谱法测量。 2-粘蛋白表征：粘蛋白分子的分离及其大分子特性的表征， 包括分子量（MW），流体动力大小（RG），电荷状态，是必要的关键参数 项目进行的计算。分子量和大小，低聚状态，形状（紧凑与线性）， 充电状态将通过GPC，折射率，多角度激光散射（购物中心）评估 琼脂糖凝胶电泳和速率区域超速离心后，然后进行电子显微镜检查。 3-粘蛋白/粘液蛋白质组：基于质谱的蛋白质组学表征培养物和， 将确定腺体分泌物以及包含粘蛋白相互作用蛋白质组的蛋白质。