Tools and Strategies to Match Peptide-Ligand Receptor Pairs
Tools and Strategies to Match Peptide-Ligand Receptor Pairs
复制标题
DOI：
10.1105/tpc.113.120071
复制
发表时间：
2014-05-01
期刊：
影响因子：
11.6
通讯作者：
Felix, Georg
中科院分区：
文献类型：
--
作者：
Butenko, Melinka A.;Wildhagen, Mari;Felix, Georg
肽信号已成为植物细胞间通讯的一类重要调节因子。具有保守的C-末端富含Pro基序的几个小的分泌蛋白家族已被鉴定为拟南芥中的功能肽信号。推测这些蛋白质经过蛋白水解修剪并经历翻译后修饰，例如Pro残基的羟基化和糖基化，以形成成熟的生物活性信号。这些配体与其各自受体的鉴定和匹配仍然是一个主要挑战，因为编码它们的基因通常显示出冗余和低表达，仅限于少数细胞或特定发育阶段。为了克服这些困难，我们建议使用异位表达的受体基因在合适的植物细胞，如本氏烟草的受体输出测定和结合研究中测试配体候选人。作为一个例子，我们使用已知的IDA肽HAE/HSL 2受体信号传导系统来调节花器官分化。我们证明，氧化爆发反应可以作为读出受体激活合成肽，一种新的，高灵敏度，非放射性标记的方法可以用来揭示肽活性和受体亲和力之间的直接相关性。我们认为，这些方法将在植物配体-受体研究领域具有广泛的价值。
Peptide signals have emerged as an important class of regulators in cell-to-cell communication in plants. Several families of small, secreted proteins with a conserved C-terminal Pro-rich motif have been identified as functional peptide signals in Arabidopsis thaliana. These proteins are presumed to be trimmed proteolytically and undergo posttranslational modifications, such as hydroxylation of Pro residues and glycosylation, to form mature, bioactive signals. Identification and matching of such ligands with their respective receptors remains a major challenge since the genes encoding them often show redundancy and low expression restricted to a few cells or particular developmental stages. To overcome these difficulties, we propose the use of ectopic expression of receptor genes in suitable plant cells like Nicotiana benthamiana for testing ligand candidates in receptor output assays and in binding studies. As an example, we used the IDA peptide HAE/HSL2 receptor signaling system known to regulate floral organ abscission. We demonstrate that the oxidative burst response can be employed as readout for receptor activation by synthetic peptides and that a new, highly sensitive, nonradioactive labeling approach can be used to reveal a direct correlation between peptide activity and receptor affinity. We suggest that these approaches will be of broad value for the field of ligand-receptor studies in plants.