Identification of global inhibitors of cellular glycosylation.
Identification of global inhibitors of cellular glycosylation.
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DOI：
10.1038/s41467-023-36598-7
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发表时间：
2023-02-20
影响因子：
16.6
通讯作者：
Narimatsu, Yoshiki
中科院分区：
文献类型：
--
作者：
Sorensen, Daniel Madriz;Bull, Christian;Madsen, Thomas D.;Lira-Navarrete, Erandi;Clausen, Thomas Mandel;Clark, Alex E.;Garretson, Aaron F.;Karlsson, Richard;Pijnenborg, Johan F. A.;Yin, Xin;Miller, Rebecca L.;Chanda, Sumit K.;Boltje, Thomas J.;Schjoldager, Katrine T.;Vakhrushev, Sergey Y.;Halim, Adnan;Esko, Jeffrey D.;Carlin, Aaron F.;Hurtado-Guerrero, Ramon;Weigert, Roberto;Clausen, Henrik;Narimatsu, Yoshiki
糖基化酶的小分子抑制剂是用于剖析聚糖功能和潜在药物候选物的有价值的工具。糖基转移酶抑制剂的筛选主要通过体外酶测定进行，难以将候选物转移到细胞和动物中。在这里，我们通过使用表达定制的报告糖蛋白的糖工程化细胞采用基于细胞的筛选测定来规避这一点。我们专注于GalNAc型O-糖基化，并选择选择性糖基化内吞低密度脂蛋白受体（LDLR）相关蛋白的GalNAc-T11同工酶作为靶标。我们的有限的小分子化合物库的筛选没有鉴定GalNAc-T11的选择性抑制剂，然而，我们鉴定了两种广泛抑制高尔基体定位的糖基化过程的化合物。这些化合物介导高尔基体系统的可逆片段化而不影响分泌。我们展示了这些抑制剂如何用于操纵细胞中的糖基化，以诱导截短O-聚糖的表达并增强癌症特异性Tn糖蛋白抗体的结合，并抑制硫酸乙酰肝素的表达以及SARS-CoV-2的结合和感染。在这里，作者发现了抑制细胞高尔基体中发生的糖基化过程的小分子。这些分子可逆地抑制精细聚糖结构的形成，而不影响糖蛋白的分泌。
Small molecule inhibitors of glycosylation enzymes are valuable tools for dissecting glycan functions and potential drug candidates. Screening for inhibitors of glycosyltransferases are mainly performed by in vitro enzyme assays with difficulties moving candidates to cells and animals. Here, we circumvent this by employing a cell-based screening assay using glycoengineered cells expressing tailored reporter glycoproteins. We focused on GalNAc-type O-glycosylation and selected the GalNAc-T11 isoenzyme that selectively glycosylates endocytic low-density lipoprotein receptor (LDLR)-related proteins as targets. Our screen of a limited small molecule compound library did not identify selective inhibitors of GalNAc-T11, however, we identify two compounds that broadly inhibited Golgi-localized glycosylation processes. These compounds mediate the reversible fragmentation of the Golgi system without affecting secretion. We demonstrate how these inhibitors can be used to manipulate glycosylation in cells to induce expression of truncated O-glycans and augment binding of cancer-specific Tn-glycoprotein antibodies and to inhibit expression of heparan sulfate and binding and infection of SARS-CoV-2. Here, the authors discover small molecules that inhibit glycosylation processes that occur in the Golgi apparatus of cells. The molecules reversibly inhibit formation of elaborate glycan structures without affecting secretion of glycoproteins.
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DOI：
10.1111/tra.12019
发表时间：
2013-01
期刊：
Traffic (Copenhagen, Denmark)
影响因子：
--
作者：
Bachert C;Linstedt AD
通讯作者：
Linstedt AD
影响因子：
16.6
作者：
Beahm, Brendan J.;Dehnert, Karen W.;Derr, Nicolas L.;Kuhn, Joachim;Eberhart, Johann K.;Spillmann, Dorothe;Amacher, Sharon L.;Bertozzi, Carolyn R.
通讯作者：
Bertozzi, Carolyn R.
影响因子：
15
作者：
Agarwal, Kavita;Kaul, Rachna;Sampathkumar, Srinivasa-Gopalan
通讯作者：
Sampathkumar, Srinivasa-Gopalan
影响因子：
48
作者：
Chen, Yen-Hsi;Narimatsu, Yoshiki;Yang, Zhang
通讯作者：
Yang, Zhang
影响因子：
6.8
作者：
de las Rivas, Matilde;Lira-Navarrete, Erandi;Hurtado-Guerrero, Ramon
通讯作者：
Hurtado-Guerrero, Ramon