Label-Free Quantification of Small-Molecule Binding to Membrane Proteins on Single Cells by Tracking Nanometer-Scale Cellular Membrane Deformation
| Autor: | Shaopeng Wang, Ashley Hunt, Hui Yu, Yunze Yang, Wenwen Jing, Hong Yuan Chen, Nongjian Tao, Fenni Zhang |
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| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Molecular binding Drug Evaluation Preclinical General Physics and Astronomy Plasma protein binding Article Cell Line Cell membrane Small Molecule Libraries 03 medical and health sciences Single-cell analysis Lab-On-A-Chip Devices medicine Humans General Materials Science Ion channel Chemistry Cell Membrane General Engineering Membrane Proteins Equipment Design Biomechanical Phenomena Kinetics 030104 developmental biology Membrane medicine.anatomical_structure Membrane protein Biophysics Small molecule binding Single-Cell Analysis Protein Binding |
| Zdroj: | ACS nano. 12(2) |
| ISSN: | 1936-086X |
| Popis: | Measuring molecular binding to membrane proteins is critical for understanding cellular functions, validating biomarkers, and screening drugs. Despite the importance, developing such a capability has been a difficult challenge, especially for small-molecule binding to membrane proteins in their native cellular environment. Here we show that the binding of both large and small molecules to membrane proteins can be quantified on single cells by trapping single cells with a microfluidic device and detecting binding-induced cellular membrane deformation on the nanometer scale with label-free optical imaging. We develop a thermodynamic model to describe the binding-induced membrane deformation, validate the model by examining the dependence of membrane deformation on cell stiffness, membrane protein expression level, and binding affinity, and study four major types of membrane proteins, including glycoproteins, ion channels, G-protein coupled receptors, and tyrosine kinase receptors. The single-cell detection capability reveals the importance of local membrane environment on molecular binding and variability in the binding kinetics of different cell lines and heterogeneity of different cells within the same cell line. |
| Databáze: | OpenAIRE |
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