New assay for screening membrane-permeabilising peptides and proteins published in Angewandte Chemie International Edition

Researchers from the ESMB have developed a single-molecule liposome assay that promises to transform the study of membrane disruption by proteins and peptides. Their innovative method, funded by the Leverhulme Trust, addresses longstanding challenges in membrane-related research across antimicrobial activity, toxicity, and disease pathology.

Membrane integrity is crucial for cellular health, playing a vital role in processes such as ion transport and signal transmission. Disruption of these membranes is linked to various disorders, including Alzheimer's and Parkinson’s disease, and certain cancers. Traditional techniques for examining the membrane-disrupting effects of proteins and peptides can be challenging and often require larger quantities, which are costly and difficult to produce.

Their new approach uses single-molecule confocal microscopy with fast-flow microfluidics to detect membrane disruption with minimal quantities of proteins and peptides. This method provides unparalleled sensitivity, allowing for the observation of individual molecular events without the need for ensemble averaging. This means researchers can capture molecular heterogeneity and subtle variations that are often lost in bulk measurements. The technology is scalable and supports high-throughput parallelisation, making it suitable for extensive applications in chemistry, biophysics, and medicine.

 

Professor Mathew Horrocks, senior author, says “We’ve traditionally used single-molecule approaches to simply look at biological systems, and so it’s exciting that we’ve now developed a method that can measure the function of biomolecules. We anticipate that our technique will streamline the development of new therapeutics and enhance our understanding of membrane dynamics in health and disease.” 

Dr Dan Edwards: “In this work, we have a designed a novel assay for the rapid determination of membrane permeabilization that can act a biological mimic to study a huge variety of anti-microbial, channel-forming, and ion-transporting mechanisms. Our assay utilises single-molecule confocal microscopy equipped with a microfluidic device which affords outstanding limits of detection and minimises the amount of material required to study the activity of an analyte.”

Link to article: https://doi.org/10.1002/anie.202503678