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Defining proteoform-specific interactions for drug targeting in a native cell signalling environment
Corinne A Lutomski1,2, Jack L Bennett1,2, Tarick J El-Baba1,2
1Physical and Theoretical Chemistry Laboratory, Department of Chemistry, University of Oxford, Oxford, UK.
Nature Chemistry
|January 13, 2025
Summary
This study reveals new methods to analyze protein modifications in native membranes, uncovering details about rhodopsin proteoforms and off-target drug interactions with retinal proteins like PDE6.
Area of Science:
- Biochemistry
- Proteomics
- Pharmacology
Background:
- Understanding membrane protein-ligand interactions is crucial for drug discovery.
- Existing cell-based assays often overlook protein modifications.
- Native lipid bilayer environments are key to protein function.
Purpose of the Study:
- To develop a method for analyzing proteoforms and their interactions in native membrane environments.
- To investigate rhodopsin proteoforms and their modifications.
- To characterize off-target drug binding to retinal proteins.
Main Methods:
- Infrared irradiation and mass spectrometry for releasing membrane proteins.
- Infrared multiphoton dissociation for proteoform sequencing.
- Characterization of off-target drug binding to phosphodiesterase 6 (PDE6).
Main Results:
- Successfully released and sequenced individual retina proteoforms, including rhodopsin.
- Identified distinct rhodopsin proteoforms, localized palmitoylations, and a Gβγ proteoform.
- Determined differential off-target binding of vardenafil and sildenafil to PDE6, with a preference for lipidated G protein proteoforms.
Conclusions:
- The study presents a novel approach for probing proteoform-ligand interactions in native membrane settings.
- Findings provide insights into rhodopsin heterogeneity and G protein lipid modifications.
- Characterization of drug off-target effects on retinal proteins offers valuable information for drug safety.
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