Department of Cellular and Molecular Medicine, University of California, San Diego, CA, USA.
Department of Clinical Sciences, Division of Infection Medicine, BMC, Lund University, Lund, Sweden.
Methods Mol Biol. 2023;2674:285-293. doi: 10.1007/978-1-0716-3243-7_19.
Vascular dysfunction is a hallmark of systemic inflammatory responses such as bacterial sepsis. The luminal surface of the blood vessels is coated with a dense layer of glycans and proteoglycans, collectively known as the glycocalyx. Surface associated glycoproteins of endothelial origin, or derived from pericytes, intravascular leukocytes, and plasma, are other important components of the glycocalyx, constituting a vascular cell surface proteome that is dynamic, tissue-specific, and sensitive to changes in vascular homeostasis, blood infection, and inflammation. Here, we describe an experimental protocol to chemically tag and quantify the vascular cell surface proteome in murine models of bacteremia, in a time-resolved and organ-specific manner. This method facilitates the identification of markers of vascular activation and provides a molecular framework to understand the contribution of vascular dysfunction to the organ pathology of systemic inflammation.
血管功能障碍是全身炎症反应(如细菌性败血症)的标志之一。血管的腔面覆盖着一层密集的糖链和糖胺聚糖,统称为糖萼。内皮来源或来源于周细胞、血管内白细胞和血浆的表面相关糖蛋白是糖萼的另一个重要组成部分,构成了一个血管细胞表面蛋白质组,它是动态的、组织特异性的,并对血管内稳态、血液感染和炎症的变化敏感。在这里,我们描述了一种实验方案,用于以时间分辨和器官特异性的方式,在细菌血症的小鼠模型中化学标记和定量血管细胞表面蛋白质组。该方法有助于鉴定血管激活的标志物,并为理解血管功能障碍对全身炎症的器官病理学的贡献提供了分子框架。