Core D: Proteomics Core Facility

摘要 核心D蛋白质组学核心设施将支持拟议的项目，以解决 蛋白质糖基化和糖蛋白重塑调节凝血和炎症的程序 败血症的症状。核心D将以无偏见的全糖蛋白组学和定向糖蛋白组学提供血液蛋白质组分析 识别血浆蛋白质组各种特殊变化的方法。血浆蛋白质组分析 将首先获得野生型小鼠和产仔未感染的血浆样本并进行比较 具有项目中指出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定量改造和 由一种新发现的分泌型糖蛋白衰老和活性机制调节丰度和活性 营业额。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细胞内凝集素受体。核心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 肺炎链球菌(SPN)、鼠伤寒沙门氏菌(ST)、霍乱沙门氏菌 (SC)和大肠杆菌(EC)。蛋白质组和糖蛋白质组的特定功能的变化将是 对这些脓毒症模型进行分析，并与可能发生的蛋白质组和糖蛋白质组变化进行比较 全身性炎症反应综合征(SIRS)模型。总血浆蛋白质组研究也将 在健康的人类志愿者以及被诊断为全身炎症反应综合征或由以下原因引起的败血症的人中进行 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 通过鉴定具有以下特性的硫酸乙酰肝素蛋白多糖(HSPG)有助于实现项目研究目标 排入脓毒症的血液中，并将进一步鉴定和量化与 循环和脱落热休克蛋白的糖胺多聚糖。Core D将进一步使用基于活动的蛋白质组学来 支持项目研究以确定基质金属蛋白酶参与凝血障碍、炎症、 脓毒症和全身炎症反应综合征的转归。同时，在上述分析中，核心D将在一定程度上充当 发现引擎，以确定有助于调节机制的循环血糖蛋白靶点 脓毒症的凝血障碍和炎症。总之，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，以解决该计划的中心假设 以进一步了解脓毒症的凝血障碍和炎症的发生和发展。

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.