Core D: Proteomics Core Facility

总结 核心D蛋白质组学核心设施将支持拟议的项目，以解决核心假设， 蛋白质糖基化和糖蛋白重塑调节凝血病和炎症的程序 败血症核心D将在无偏倚的总糖蛋白组学和定向糖蛋白组学中提供血液蛋白组学分析 确定血浆蛋白质组的各种特定变化的方法。血浆蛋白质组学分析 将首先获得野生型小鼠和同窝小鼠的未感染血浆样品并进行比较 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重塑的血浆蛋白， 通过新发现的分泌糖蛋白老化机制调节丰度和活性， 周转这些研究涉及接受糖苷酶抑制剂的小鼠，缺乏糖苷酶， 调节血浆糖蛋白稳态的内吞凝集素受体。核心D将进一步提供 血浆蛋白质组学分析在由不同细菌病原体感染引起的小鼠脓毒症中的应用， 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的特定特征的变化将被 在这些脓毒症模型中进行了分析，并与可能发生在 全身炎症反应综合征（SIRS）的模型。总血浆蛋白质组学研究也将 在健康的人类志愿者以及被诊断患有由以下疾病引起的SIRS或脓毒症的人中进行 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 通过鉴定硫酸乙酰肝素蛋白聚糖（HSPG），有助于实现项目研究目标， 在败血症中流入血液，并将进一步识别和量化与 循环和脱落的HSPG的糖胺聚糖。核心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， 以及脓毒症和全身炎症反应综合征的结果。在上述试验中，核心D将部分地充当 发现引擎，以确定循环血液糖蛋白的目标，有助于调节机制， 凝血病和败血症炎症。总体而言，核心D将促进实现协同增效， 研究结果，整合所有三个项目的数据，以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。

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.