Work

Simple methods to determine the dissociation constant, Kd

Year: 2024

Type: article

Abstract: The determination of the dissociation constant (K d ) is pivotal in biochemistry and pharmacology for understanding binding affinities in chemical reactions, which is crucial for drug development and comprehending biological systems. Here, we introduce a single-molecule fluorescence resonance energy transfer–based method for determining K d , alongside the conventional electrophoretic mobility shift assay method of K d , offering insights into thermodynamic interactions between proteins and substrates. The single-molecule fluorescence resonance energy transfer approach is highlighted for its ability to accurately measure binding and dissociation kinetics through fluorescence labeling and the intrinsic nature of protein-DNA interactions, representing a significant advancement in the fields of molecular biology and pharmacology.

Source: Molecules and Cells

DOI: 10.1016/j.mocell.2024.100112

Authors: Donghun Lee, Juwon Kim, Gwangrog Lee

Institution: Korea Advanced Institute of Science and Technology

Language: English

Topic: Advanced Fluorescence Microscopy Techniques

Subfield: Biophysics

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

Open Access status: gold

Estimated APC paid: $2,500

Funder: Korea Advanced Institute of Science and Technology