Key Laboratory of Ministry of Education for Protein Science, School of Life Sciences, Tsinghua University , Beijing 100084, P.R. China.
Anal Chem. 2017 Jul 5;89(13):6926-6930. doi: 10.1021/acs.analchem.7b01274. Epub 2017 Jun 22.
The induced dimerization of two distinct receptors through a heterobifunctional inducer is prevalent among all levels of cellular signaling processes, yet its complexity poses difficulty for systematic quantitative analysis. This paper first shows how to calculate the amount of any possible complex or monomer of heteroligand and two receptors present at equilibrium. The theory is subsequently applied to the determination of three independent equilibrium parameters involved in the rapamycin induced FKBP and FRB dimerization, in which all parameters were simultaneously estimated using one set of fluorescence resonance energy transfer (FRET) experiments. A MATLAB script is provided for parametric fitting.
通过杂双功能诱导剂诱导两个不同受体的二聚化在细胞信号转导过程的所有层次中都很普遍,但它的复杂性给系统的定量分析带来了困难。本文首先展示了如何计算任何可能的复合物或单体的量,以及在平衡状态下存在的两种受体。随后,该理论被应用于确定雷帕霉素诱导的 FKBP 和 FRB 二聚化过程中涉及的三个独立平衡参数,其中所有参数都是使用一组荧光共振能量转移 (FRET) 实验同时估计的。提供了一个 MATLAB 脚本用于参数拟合。