Core D: Proteomics Core Facility

核心 D：蛋白质组学核心设施

• 批准号: 9072752
• 负责人: Jeffrey W Smith
• 金额: $ 37.95万
• 依托单位: SANFORD BURNHAM PREBYS MEDICAL DISCOVERY INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-07-15 至 2021-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9072752
• 关键词: Active Sites; Address; Affinity Chromatography; Aging; Binding Proteins; Biological Assay; Blood; Blood Circulation; Blood Coagulation Disorders; California; Cell surface; Censuses; Complex; Computer software; Core Facility; Data; Data Analyses; Diagnosis; Dissociation; Electron Transport; Endothelial Cells; Escherichia coli; Genetic; Glycoproteins; Glycosaminoglycans; Glycoside Hydrolases; Goals; Healthcare; Heparan Sulfate Proteoglycan; Homeostasis; Hospitals; Human; Human Volunteers; Immunoprecipitation; Individual; Infection; Inflammation; Investigation; Lectin; Lectin Receptors; Mass Spectrum Analysis; Matrix Metalloproteinase Inhibitor; Matrix Metalloproteinases; Mediating; Medical Research; Microbe; Modeling; Mus; N acetylglucosaminidase; Neuraminidase; Outcome; Pathogenesis; Patients; Plasma; Plasma Proteins; Post Translational Modification Analysis; Preparation; Protein Glycosylation; Proteins; Proteome; Proteomics; Regulation; Research; Research Institute; Research Project Grants; Running; Salmonella enterica; Salmonella typhimurium; Sampling; Sepsis; Sepsis Syndrome; Streptavidin; Streptococcus pneumoniae; Surface; Syndrome; Technology; Wild Type Mouse; activity-based protein profiling; base; beta-Galactosidase; cost; glycoproteomics; healthy volunteer; inhibitor/antagonist; instrumentation; mass spectrometer; mouse model; pathogen; programs; protective effect; receptor; research study; senescence; septic; therapeutic target; trauma centers

SUMMARY The Core D proteomics core facility will support the proposed project to address the central hypothesis of the program that Protein glycosylation and glycoprotein remodeling modulate the coagulopathy and inflammation of sepsis. Core D will provide blood proteomic analyses in unbiased total and directed glycoproteomic approaches to identify various and specific changes to blood plasma proteomes. Plasma proteomic analyses will be first obtained and compared among uninfected plasma samples of wild-type mice and littermates bearing specific genetic deficiency states as indicated in the projects. Core D will contribute to achieving relevant project research aims in part by identifying and quantifying plasma proteins that are remodeled and regulated in abundance and activity by a newly discovered mechanism of secreted glycoprotein aging and turnover. These studies involve mice receiving glycosidase inhibitors, lacking glycosidases, and lacking endocytic lectin receptors that regulate blood plasma glycoprotein homeostasis. Core D will further provide plasma proteomic analyses in mouse sepsis arising from infections with different bacterial pathogens including Streptococcus pneumoniae (SPN), Salmonella enterica Typhimurium (ST), Salmonella enterica Choleraesuis (SC) and Escherichia coli (EC). Changes to the proteome and to specific features of the glycoproteome will be analyzed in these sepsis models and compared with proteomic and glycoproteomic changes that may occur in a model of the Systemic Inflammatory Response Syndrome (SIRS). Total plasma proteomic studies will also be performed among healthy human volunteers as well as those diagnosed with SIRS or sepsis caused by Gram-negative (mostly EC) or Gram-positive (including pneumococcal) bacterial pathogens. Core D will also contribute to achieving project research aims by identifying heparan sulfate proteoglycans (HSPGs) that are shed into the bloodstream in sepsis, and will further identify and quantify proteins that are bound to the glycosaminoglycans of circulating and shed HSPGs. Core D will further use activity-based proteomics to support project research to identify matrix metalloproteinases participating in the coagulopathy, inflammation, and outcomes of sepsis and SIRS. Together and in the aforementioned assays, Core D will act in part as a discovery engine to identify circulating blood glycoprotein targets that contribute to mechanisms modulating the coagulopathy and inflammation of sepsis. Altogether Core D will promote the attainment of synergistic research findings that integrate data from all three projects in addressing the central hypothesis of the program to further understand the onset and progression of the coagulopathy and inflammation of sepsis.

概括 Core D 蛋白质组学核心设施将支持拟议的项目，以解决以下问题的中心假设： 蛋白质糖基化和糖蛋白重塑调节凝血病和炎症的程序 败血症。 Core D 将提供无偏见的总体和定向糖蛋白质组学血液蛋白质组学分析 识别血浆蛋白质组的各种特定变化的方法。血浆蛋白质组分析 将首先获得并比较野生型小鼠和同窝小鼠的未感染血浆样本 具有项目中所示的特定遗传缺陷状态。核心D将有助于实现 相关项目研究的部分目的是通过识别和量化被重塑和 通过新发现的分泌糖蛋白老化机制调节其丰度和活性 周转。这些研究涉及接受糖苷酶抑制剂、缺乏糖苷酶和缺乏糖苷酶的小鼠。 调节血浆糖蛋白稳态的内吞凝集素受体。 Core D将进一步提供 不同细菌病原体感染引起的小鼠脓毒症的血浆蛋白质组学分析，包括 肺炎链球菌 (SPN)、鼠伤寒沙门氏菌 (ST)、猪霍乱沙门氏菌 (SC) 和大肠杆菌 (EC)。蛋白质组和糖蛋白质组特定特征的变化将是 在这些脓毒症模型中进行分析，并与可能发生的蛋白质组和糖蛋白质组变化进行比较 全身炎症反应综合征（SIRS）模型。总血浆蛋白质组学研究也将 在健康人类志愿者以及被诊断患有 SIRS 或脓毒症的志愿者中进行 革兰氏阴性（主要是 EC）或革兰氏阳性（包括肺炎球菌）细菌病原体。核心D也将 通过鉴定硫酸乙酰肝素蛋白聚糖 (HSPG) 来帮助实现项目研究目标 在脓毒症中流入血液，并将进一步识别和量化与 循环和脱落的 HSPG 的糖胺聚糖。 Core D 将进一步使用基于活性的蛋白质组学 支持项目研究，以确定参与凝血病、炎症、 以及脓毒症和 SIRS 的结果。在上述测定中，Core D 将部分充当 发现引擎，用于识别有助于调节机制的循环血糖蛋白靶点 凝血障碍和脓毒症炎症。总而言之，核心 D 将促进协同效应的实现 研究结果整合了所有三个项目的数据来解决该计划的中心假设 进一步了解脓毒症凝血障碍和炎症的发生和进展。