Mapping glycan-mediated galectin-3 interactions by live cell proximity labeling.
Mapping glycan-mediated galectin-3 interactions by live cell proximity labeling.
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DOI：
10.1073/pnas.2009206117
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发表时间：
2020-11-03
影响因子：
11.1
通讯作者：
Huang ML
中科院分区：
文献类型：
--
作者：
Joeh E;O'Leary T;Li W;Hawkins R;Hung JR;Parker CG;Huang ML
由于单个糖链结合蛋白(GBP)之间的弱相互作用，如Galectin-3和多糖之间的相互作用，使得能够在活细胞中直接询问这些相互作用的策略仍然有限。因此，在生理上与Galectin-3结合相关的糖链和糖蛋白配体的描述还不够充分。在这里，我们使用了一种邻近标记方法，该方法催化标记Galectin-3的相互作用，并在活细胞中鉴定其相关的糖蛋白和糖蛋白拮抗剂受体。这项研究表明，邻近标记是在活细胞中定位GBP复合体的强大工具，当与化学抑制剂结合时，它可以区分蛋白质-蛋白质和蛋白质-多糖相互作用。Galectin-3是一种糖链结合蛋白，能与β-半乳糖苷糖链结构结合，协调多种重要的生物学事件，包括激活肝星状细胞和调节免疫反应。虽然与Galectin-3结合所需的糖链表位早已被阐明，但承载这些糖链特征的细胞糖蛋白仍然未知。鉴于三维(3D)多糖排列在决定GBP相互作用中的重要性，允许在活细胞中识别GBP受体的策略具有显著的优势，因为在活细胞中保留了天然的糖提呈和糖蛋白表达。在这里，我们描述了一种邻近标记方法和定量质谱学的集成，以定位活的人肝星状细胞和外周血单核细胞中Galectin-3的糖蛋白和糖蛋白的相互作用。了解糖蛋白的特性和确定糖聚糖的结构将有助于设计和开发选择性疗法，以减轻Galectin-3介导的生物事件。
Because of the weak interactions between individual glycan-binding proteins (GBPs), such as galectin-3, and glycans, strategies that enable the direct interrogation of these interactions in living cells remain limited. Thus, the glycan and glycoprotein ligands that are physiologically relevant for galectin-3 binding are insufficiently described. Here we used a proximity labeling approach that catalytically tags interactors for galectin-3 and identified its pertinent glycan and glycoprotein counter-receptors in live cells. This study demonstrates that proximity labeling is a powerful tool for mapping GBP complexes in living cells, and when coupled with chemical inhibitors, it can discriminate between protein–protein and protein–glycan interactions. Galectin-3 is a glycan-binding protein (GBP) that binds β-galactoside glycan structures to orchestrate a variety of important biological events, including the activation of hepatic stellate cells and regulation of immune responses. While the requisite glycan epitopes needed to bind galectin-3 have long been elucidated, the cellular glycoproteins that bear these glycan signatures remain unknown. Given the importance of the three-dimensional (3D) arrangement of glycans in dictating GBP interactions, strategies that allow the identification of GBP receptors in live cells, where the native glycan presentation and glycoprotein expression are preserved, have significant advantages over static and artificial systems. Here we describe the integration of a proximity labeling method and quantitative mass spectrometry to map the glycan and glycoprotein interactors for galectin-3 in live human hepatic stellate cells and peripheral blood mononuclear cells. Understanding the identity of the glycoproteins and defining the structures of the glycans will empower efforts to design and develop selective therapeutics to mitigate galectin-3–mediated biological events.
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影响因子：
14.8
作者：
Hung V;Udeshi ND;Lam SS;Loh KH;Cox KJ;Pedram K;Carr SA;Ting AY
通讯作者：
Ting AY
影响因子：
3
作者：
Haudek, Kevin C.;Spronk, Kimberly J.;Voss, Patricia G.;Patterson, Ronald J.;Wang, John L.;Arnoys, Eric J.
通讯作者：
Arnoys, Eric J.
DOI：
10.1074/jbc.c112.358002
发表时间：
2012-06-22
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Lepur A;Salomonsson E;Nilsson UJ;Leffler H
通讯作者：
Leffler H
影响因子：
56.9
作者：
Brightbill, HD;Libraty, DH;Modlin, RL
通讯作者：
Modlin, RL
影响因子：
56.9
作者：
Aliprantis, AO;Yang, RB;Zychlinsky, A
通讯作者：
Zychlinsky, A