Miyashita Masahiro, Miyagawa Hisashi
Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Kyoto, Japan.
Comb Chem High Throughput Screen. 2008 Sep;11(8):647-52. doi: 10.2174/138620708785739970.
As plants lack a circulatory system and adaptive immune system, they have evolved their own defense systems distinct from animals, in which each plant cell is capable of defending itself from pathogens. Plants induce a number of defense responses, which are triggered by a variety of molecules derived from pathogenic microorganisms, referred to as microbe-associated molecular patterns (MAMPs), including peptides, proteins, lipopolysaccharide, beta-glucan, chitin, and ergosterol. The interaction between plants and chemicals in the context of plant defense represents a "natural" and simple model for chemogenomics, at the intersection between chemical and biological diversities. For protection of crop plants from diseases, it has been shown to be effective to stimulate the plant immunity by chemical compounds, the so-called "plant defense activators". Combinatorial chemistry techniques can be applied to the search for novel plant defense activators, but it is essential to establish an efficient and reliable screening system suitable for library screening. For studies of the plant immune system, it is difficult to use isolated proteins as biological targets because the receptors for MAMP recognition are largely unknown and even the receptors identified so far are transmembrane proteins. Therefore, screening for novel peptides acting on MAMP receptors from combinatorial libraries must rely on a solution-phase assay using cells as the biological targets. In this review, we introduce the cell-based lawn format assay for identification of peptides acting as plant defense activators from combinatorial peptide libraries. The requirements and limitations in constructing the screening system using combinatorial libraries in the studies of plant sciences are also discussed.
由于植物缺乏循环系统和适应性免疫系统,它们进化出了与动物不同的自身防御系统,其中每个植物细胞都能够抵御病原体。植物会引发多种防御反应,这些反应由多种源自病原微生物的分子触发,这些分子被称为微生物相关分子模式(MAMPs),包括肽、蛋白质、脂多糖、β-葡聚糖、几丁质和麦角固醇。在植物防御的背景下,植物与化学物质之间的相互作用代表了化学基因组学的一个“自然”且简单的模型,处于化学和生物多样性的交叉点。为了保护农作物免受病害侵袭,已证明通过化合物(即所谓的“植物防御激活剂”)刺激植物免疫是有效的。组合化学技术可应用于寻找新型植物防御激活剂,但建立一个适用于文库筛选的高效可靠的筛选系统至关重要。对于植物免疫系统的研究,由于MAMP识别受体大多未知,甚至到目前为止所鉴定的受体都是跨膜蛋白,因此很难将分离的蛋白质用作生物学靶点。所以,从组合文库中筛选作用于MAMP受体的新型肽必须依赖以细胞作为生物学靶点的液相分析。在这篇综述中,我们介绍了基于细胞的平板法,用于从组合肽文库中鉴定作为植物防御激活剂的肽。还讨论了在植物科学研究中使用组合文库构建筛选系统的要求和局限性。