Unveiling the multi-step solubilization mechanism of sub-micron size vesicles by detergents

Dalgarno, P. A. et al. (2019) Unveiling the multi-step solubilization mechanism of sub-micron size vesicles by detergents. Scientific Reports, 9, 12897. (doi: 10.1038/s41598-019-49210-0) (PMID:31501469) (PMCID:PMC6733941)

[img] Text
196713.pdf - Published Version
Available under License Creative Commons Attribution.



The solubilization of membranes by detergents is critical for many technological applications and has become widely used in biochemistry research to induce cell rupture, extract cell constituents, and to purify, reconstitute and crystallize membrane proteins. The thermodynamic details of solubilization have been extensively investigated, but the kinetic aspects remain poorly understood. Here we used a combination of single-vesicle Förster resonance energy transfer (svFRET), fluorescence correlation spectroscopy and quartz-crystal microbalance with dissipation monitoring to access the real-time kinetics and elementary solubilization steps of sub-micron sized vesicles, which are inaccessible by conventional diffraction-limited optical methods. Real-time injection of a non-ionic detergent, Triton X, induced biphasic solubilization kinetics of surface-immobilized vesicles labelled with the Dil/DiD FRET pair. The nanoscale sensitivity accessible by svFRET allowed us to unambiguously assign each kinetic step to distortions of the vesicle structure comprising an initial fast vesicle-swelling event followed by slow lipid loss and micellization. We expect the svFRET platform to be applicable beyond the sub-micron sizes studied here and become a unique tool to unravel the complex kinetics of detergent-lipid interactions.

Item Type:Articles
Additional Information:The authors thank Dr. Arvydas Ruseckas for helping with fluorescence lifetime measurements and the EPSRC (EP/P030017) for support.
Glasgow Author(s) Enlighten ID:Hedley, Dr Gordon
Authors: Dalgarno, P. A., Juan-Colás, J., Hedley, G. J., Piñeiro, L., Novo, M., Perez-Gonzalez, C., Samuel, I. D. W., Leake, M. C., Johnson, S., Al-Soufi, W., Penedo, J. C., and Quinn, S. D.
College/School:College of Science and Engineering > School of Chemistry
Journal Name:Scientific Reports
Publisher:Nature Research
ISSN (Online):2045-2322
Copyright Holders:Copyright © 2019 The Authors
First Published:First published in Scientific Reports 9: 12897
Publisher Policy:Reproduced under a Creative Commons License

University Staff: Request a correction | Enlighten Editors: Update this record