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- Title
Unveiling the multi-step solubilization mechanism of sub-micron size vesicles by detergents.
- Authors
Dalgarno, Paul A.; Juan-Colás, José; Hedley, Gordon J.; Piñeiro, Lucas; Novo, Mercedes; Perez-Gonzalez, Cibran; Samuel, Ifor D. W.; Leake, Mark C.; Johnson, Steven; Al-Soufi, Wajih; Penedo, J. Carlos; Quinn, Steven D.
- Abstract
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.
- Subjects
SOLUBILIZATION; DETERGENTS; MEMBRANE proteins; SPECTROMETRY; QUARTZ crystal microbalances
- Publication
Scientific Reports, 2019, Vol 9, Issue 1, pN.PAG
- ISSN
2045-2322
- Publication type
Article
- DOI
10.1038/s41598-019-49210-0