Bridging the Glycome and Proteome with Chemical Biology

用化学生物学连接糖组和蛋白质组

基本信息

批准号：
10798880
负责人：
Mia L Huang
金额：
$ 8.65万
依托单位：
SCRIPPS RESEARCH INSTITUTE, THE
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-06-01 至 2026-04-30
项目状态：
未结题

项目摘要

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.

项目概要怀孕期间，胎盘的正常发育对于维持胎儿的健康至关重要。母亲;然而，我们仍然不清楚胎盘的某些成分在正常情况下是如何运作的。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，并且了解这个过程，我们将重点关注呈现在称为细胞表面的屏障外部的蛋白质蛋白质（CSP）。 CSP 是细胞间相互作用的调节剂，帮助细胞对其环境做出反应，使它们具有重要的生物学意义和相关的药物靶点。然而，CSP 的研究具有挑战性，因为它们丰度较低，并被称为聚糖的糖分子覆盖。这些聚糖高度无法预测的可变结构，需要实验证据来确定其组成。一这些聚糖最重要的功能之一是与聚糖结合蛋白（GBP）相互作用。英镑绑定到细胞表面的聚糖，导致细胞行为的下游变化。 GBP 与半乳糖凝集素 3 一样，已被已被证明对于正常胎儿发育和胎盘细胞融合是必需的，但尚未发现其机制行动具有挑战性，因为聚糖和 GBP 之间的弱且动态的相互作用并不容易捕获。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活细胞中的 CSP 和 galectin-3 相互作用物。我们将把这些方法应用到胎盘细胞融合模型中了解导致成功形成障碍的因素。一旦关键的 CSP 和半乳糖凝集素 3 相互作用因子被一旦发现，我们将进行遗传和生化实验来详细确定它们的功能。我们将绘制细胞内允许融合的信号通路，以及允许融合的聚糖的组成英镑互动。