Uebler Susanne, Dresselhaus Thomas
*Cell Biology and Plant Biochemistry, Biochemie-Zentrum Regensburg, University of Regensburg, 93053 Regensburg, Germany.
Biochem Soc Trans. 2014 Apr;42(2):395-400. doi: 10.1042/BST20130251.
Cell-cell communication during development and reproduction in plants depends largely on a few phytohormones and many diverse classes of polymorphic secreted peptides. The peptide ligands are bound at the cell surface of target cells by their membranous interaction partners representing, in most cases, either receptor-like kinases or ion channels. Although knowledge of both the extracellular ligand and its corresponding receptor(s) is necessary to describe the downstream signalling pathway(s), to date only a few ligand-receptor pairs have been identified. Several methods, such as affinity purification and yeast two-hybrid screens, have been used very successfully to elucidate interactions between soluble proteins, but most of these methods cannot be applied to membranous proteins. Experimental obstacles such as low concentration and poor solubility of membrane receptors, as well as instable transient interactions, often hamper the use of these 'classical' approaches. However, over the last few years, a lot of progress has been made to overcome these problems by combining classical techniques with new methodologies. In the present article, we review the most promising recent methods in identifying cell-surface receptor interactions, with an emphasis on success stories outside the field of plant research.
植物发育和繁殖过程中的细胞间通讯很大程度上依赖于少数植物激素和许多不同类别的多态性分泌肽。肽配体在靶细胞的细胞表面通过其膜相互作用伙伴结合,在大多数情况下,这些伙伴代表受体样激酶或离子通道。虽然要描述下游信号通路,细胞外配体及其相应受体的知识都必不可少,但迄今为止,仅鉴定出少数配体-受体对。几种方法,如亲和纯化和酵母双杂交筛选,已非常成功地用于阐明可溶性蛋白质之间的相互作用,但这些方法大多不适用于膜蛋白。诸如膜受体浓度低、溶解性差以及不稳定的瞬时相互作用等实验障碍,常常阻碍这些“经典”方法的使用。然而,在过去几年中,通过将经典技术与新方法相结合,在克服这些问题方面取得了很大进展。在本文中我们综述了鉴定细胞表面受体相互作用的最有前景的最新方法,重点介绍植物研究领域之外的成功案例。