Ultrasensitive Label-Free Detection of Protein-Membrane Interaction Exemplified by Toxin-Liposome Insertion.
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BORIS DOI
Date of Publication
April 14, 2022
Publication Type
Article
Division/Institute
Author
Schönfeldová, T | |
Okur, H I | |
Vezočnik, V | |
Cao, C | |
Dal Peraro, M | |
Maček, P | |
Roke, S |
Subject(s)
Series
The journal of physical chemistry letters
ISSN or ISBN (if monograph)
1948-7185
Publisher
American Chemical Society
Language
English
Publisher DOI
PubMed ID
35377651
Description
Measuring the high-affinity binding of proteins to liposome membranes remains a challenge. Here, we show an ultrasensitive and direct detection of protein binding to liposome membranes using high throughput second harmonic scattering (SHS). Perfringolysin O (PFO), a pore-forming toxin, with a highly membrane selective insertion into cholesterol-rich membranes is used. PFO inserts only into liposomes with a cholesterol concentration >30%. Twenty mole-percent cholesterol results in neither SHS-signal deviation nor pore formation as seen by cryo-electron microscopy of PFO and liposomes. PFO inserts into cholesterol-rich membranes of large unilamellar vesicles in an aqueous solution with Kd = (1.5 ± 0.2) × 10-12 M. Our results demonstrate a promising approach to probe protein-membrane interactions below sub-picomolar concentrations in a label-free and noninvasive manner on 3D systems. More importantly, the volume of protein sample is ultrasmall (<10 μL). These findings enable the detection of low-abundance proteins and their interaction with membranes.
File(s)
File | File Type | Format | Size | License | Publisher/Copright statement | Content | |
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acs.jpclett.1c04011.pdf | text | Adobe PDF | 1.53 MB | published |