Rodriguez-Furlán Cecilia, Hicks Glenn R, Norambuena Lorena
Plant Molecular Biology Laboratory, Department of Biology, University of Chile, Las Palmeras 3425, 7800003, Santiago, Chile.
Methods Mol Biol. 2014;1174:317-28. doi: 10.1007/978-1-4939-0944-5_22.
The plant endomembrane trafficking system is a highly complex set of processes. This complexity presents a challenge for its study. Classical plant genetics often struggles with loss-of-function lethality and gene redundancy. Chemical genomics allows overcoming many of these issues by using small molecules of natural or synthetic origin to inhibit specific trafficking proteins thereby affecting the processes in a tunable and reversible manner. Bioactive chemicals identified by high-throughput phenotype screens must be characterized in detail starting with understanding of the specific trafficking pathways affected. Here, we describe approaches to characterize bioactive compounds that perturb vesicle trafficking. This should equip researchers with practical knowledge on how to identify endomembrane-specific trafficking pathways that may be perturbed by specific compounds and will help to eventually identify molecular targets for these small molecules.
植物内膜运输系统是一组高度复杂的过程。这种复杂性给其研究带来了挑战。经典的植物遗传学常常面临功能丧失致死性和基因冗余的问题。化学基因组学通过使用天然或合成来源的小分子来抑制特定的运输蛋白，从而以可调节和可逆的方式影响这些过程，有助于克服其中许多问题。通过高通量表型筛选鉴定出的生物活性化学物质，必须从了解受影响的特定运输途径开始进行详细表征。在这里，我们描述了表征干扰囊泡运输的生物活性化合物的方法。这将为研究人员提供关于如何识别可能被特定化合物干扰的内膜特异性运输途径的实用知识，并最终有助于确定这些小分子的分子靶点。
