Université Paris-Saclay, INRAE, AgroParisTech, Institut Jean-Pierre Bourgin (IJPB), Versailles, France.
Biology Department, Integrated Molecular Plant Physiology Research, University of Antwerp, Antwerpen, Belgium.
Methods Mol Biol. 2024;2731:279-293. doi: 10.1007/978-1-0716-3511-7_21.
Microscale thermophoresis (MST) is a simple but powerful tool to study the in vitro interaction among biomolecules, and to quantify binding affinities. MST curves describe the change in the fluorescence level of a fluorescent target as a result of an IR-laser-induced temperature change. The degree and nature of the change in fluorescence signal depends on the size, charge, and solvation shell of the molecules, properties that change in function of the binding of a ligand to the fluorescent target.We used MST to describe the interaction between components of a regulatory module involved in plant cell wall integrity control. This module comprises the secreted peptide Rapid Alkalinization Factor 23 (RALF23) and its receptor complex consisting of the GPI-anchored receptor Lorelei-Like Glycoprotein 1 (LLG1) and a receptor kinase of the CrRLK1L family, FERONIA. Here we show how MST can also be used to study three-partner interactions.
微尺度热泳（MST）是一种简单但强大的工具，可用于研究生物分子之间的体外相互作用，并定量结合亲和力。MST 曲线描述了荧光靶标荧光强度随红外激光诱导温度变化而发生的变化。荧光信号变化的程度和性质取决于分子的大小、电荷和溶剂化壳，这些性质会随着配体与荧光靶标的结合而发生变化。我们使用 MST 来描述参与植物细胞壁完整性控制的调节模块的成分之间的相互作用。该模块包括分泌肽快速碱化因子 23（RALF23）及其由糖基磷脂酰肌醇锚定受体 Lorelei-Like Glycoprotein 1（LLG1）和 CrRLK1L 家族的受体激酶 FERONIA 组成的受体复合物。在这里，我们展示了 MST 如何也可用于研究三组分相互作用。
