Bridging the Glycome and Proteome with Chemical Biology

项目总结 在怀孕期间，胎盘的正常发育对维持胎儿的健康和 然而，我们仍然不清楚胎盘的某些成分是如何正常的 形式。我们感兴趣的是合体滋养层屏障是如何通过细胞融合形成的，并 了解这一过程，我们将集中在屏障外部的蛋白质，称为细胞表面 蛋白质(CSP)。CSP是细胞间相互作用的调节者，帮助细胞对环境做出反应，使 它们具有重要的生物学意义和相关的药物靶点。然而，CSP的研究是具有挑战性的，因为他们 都是低丰度的，覆盖着一种叫做多糖的糖分子。这些葡聚糖含量很高 无法预测的可变结构，需要实验证据来确定它们的组成。一 这些多糖最重要的功能之一是与糖结合蛋白(GBP)相互作用。Gbps绑定到 细胞表面的葡聚糖，导致下游细胞行为的变化。Gbps，像Galectin-3一样，一直是 被证明是正常胎儿发育和胎盘细胞融合所必需的，但发现了它们的机制 由于葡聚糖和GBP之间的弱而动态的相互作用并不容易 抓捕。为了克服这些限制，我们开发了两种新技术，使我们能够标记和识别 CSPs和Galectin-3在活细胞中的相互作用。我们将把这些方法应用于胎盘细胞融合的模型 了解导致成功形成障碍的因素。一旦关键的CSP和Galectin-3相互作用 发现后，我们将进行遗传和生化实验，以详细确定它们的功能。我们会 绘制细胞内允许融合的信号通路，以及允许融合的多糖的组成 用于英镑的互动。

PROJECT SUMMARY During pregnancy, the proper development of the placenta is essential to maintain the health of the fetus and mother; however, we still do not have a clear understanding of how certain components of the placenta normally form. We are interested in how the synctitiotrophoblast barrier is formed through cellular fusion, and to understand this process, we will focus on the proteins presented on the outside of the barrier called cell surface proteins (CSPs). CSPs are regulators of cell-cell interactions and help cells respond to their environment, making them biologically important and relevant drug targets. However, CSPs are challenging to study, because they are in low abundance and are covered with sugar molecules called glycans. These glycans are highly variable structures that cannot be predicted, requiring experimental evidence to ascertain their composition. One of the most important functions of these glycans is to interact with glycan-binding proteins (GBPs). GBPs bind to glycans on the cell surface, resulting in downstream changes to cell behavior. GBPs, like galectin-3, have been shown to be necessary for normal fetal development and placental cell fusion, but discovering their mechanism of action is challenging, as the weak and dynamic interactions between glycans and GBPs are not easy to capture. To overcome these limitations, we have developed two new techniques that allow us to label and identify CSPs and galectin-3 interactors in live cells. We will apply these methods to a model of placental cell fusion to understand the factors that lead to successful barrier formation. Once critical CSPs and galectin-3 interactors are uncovered, we will perform genetic and biochemical experiments to determine their functions in detail. We will map the signaling pathways within the cell that allow for fusion, as well as the composition of glycans that allow for GBP interactions.