Secramine inhibits Cdc42-dependent functions in cells and Cdc42 activation in vitro
Secramine inhibits Cdc42-dependent functions in cells and Cdc42 activation in vitro
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DOI：
10.1038/nchembio751
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发表时间：
2006-01-01
影响因子：
14.8
通讯作者：
Kirchhausen, T
中科院分区：
文献类型：
--
作者：
Henry, EP;Peterson, JR;Kirchhausen, T
受小分子研究膜传输有用性的启发，我们使用高通量合成和表型筛选发现了沙敏，这是一种通过未知机制抑制高尔基体膜传输的分子。我们在这里报道，沙明通过依赖于鸟嘌呤解离抑制剂RhoGDI的机制抑制Rho GTPase Cdc42的激活，Rho GTPase Cdc42是一种参与膜运输的蛋白质。RhoGDI结合Cdc42并拮抗其膜结合、核苷酸交换和效应物结合。在体外，沙明以rhogdi依赖的方式抑制Cdc42与膜、GTP和效应器的结合。在细胞中，沙明模拟显性负Cdc42表达对高尔基体蛋白输出和迁移细胞中高尔基体极化的影响。通过沙明抑制RhoGDI依赖性Cdc42为小分子抑制Rho gtpase提供了新的途径，为研究Cdc42、RhoGDI及其介导的细胞过程提供了新的手段。
Inspired by the usefulness of small molecules to study membrane traffic, we used high-throughput synthesis and phenotypic screening to discover secramine, a molecule that inhibits membrane traffic out of the Golgi apparatus by an unknown mechanism. We report here that secramine inhibits activation of the Rho GTPase Cdc42, a protein involved in membrane traffic, by a mechanism dependent upon the guanine dissociation inhibitor RhoGDI. RhoGDI binds Cdc42 and antagonizes its membrane association, nucleotide exchange and effector binding. In vitro, secramine inhibits Cdc42 binding to membranes, GTP and effectors in a RhoGDI-dependent manner. In cells, secramine mimics the effects of dominant-negative Cdc42 expression on protein export from the Golgi and on Golgi polarization in migrating cells. RhoGDI-dependent Cdc42 inhibition by secramine illustrates a new way to inhibit Rho GTPases with small molecules and provides a new means to study Cdc42, RhoGDI and the cellular processes they mediate.