利用 SPR 方法研究小分子-蛋白质相互作用。

Characterization of Small Molecule-Protein Interactions Using SPR Method.

机构信息

College of Horticulture, South China Agricultural University, Guangzhou, China.

Department of Molecular Biosciences and Bioengineering, University of Hawaii at Manoa, Honolulu, HI, USA.

出版信息

Methods Mol Biol. 2023;2690:149-159. doi: 10.1007/978-1-0716-3327-4_15.

DOI:10.1007/978-1-0716-3327-4_15

PMID:37450146

Abstract

Surface plasmon resonance (SPR) is an optical phenomenon being used to monitor molecular binding events. With the advantages of being label-free, real-time, and sensitive, SPR assays have become one of the most commonly used techniques to measure binding kinetics, affinity, specificity, and concentration of molecular interactions. In an SPR experiment to measure small molecule-protein interactions, the protein is immobilized on the biosensor surface, while the small molecule flows through the surface of the sensor chip. The interactions between the small molecules and proteins are monitored by subsequent changes in the refractive index and quantified with resonance units. In this chapter, we have utilized an SPR assay to study the interaction of flavonoids and the glucose-regulated protein 78. Assay steps are detailed for immobilization optimization, SPR instrument setup, operation, sample injection, and affinity data analysis.

摘要

表面等离子体共振（SPR）是一种用于监测分子结合事件的光学现象。由于具有无标记、实时和灵敏的优点，SPR 分析已成为测量分子相互作用的结合动力学、亲和力、特异性和浓度的最常用技术之一。在 SPR 实验中测量小分子-蛋白质相互作用时，将蛋白质固定在生物传感器表面，而小分子则流过传感器芯片表面。通过随后的折射率变化来监测小分子和蛋白质之间的相互作用，并用量化的共振单位来定量。在本章中，我们利用 SPR 分析来研究黄酮类化合物与葡萄糖调节蛋白 78 的相互作用。详细介绍了固定化优化、SPR 仪器设置、操作、样品注入和亲和力数据分析的步骤。