RT Journal Article
T1 Unveiling the multi-step solubilization mechanism of sub-micron size vesicles by detergents
A1 Dalgarno, Paul A.
A1 Juan-Colás, José
A1 Hedley, Gordon J.
A1 Piñeiro Maseda, Lucas
A1 Pérez González, Cibrán
A1 Samuel, Ifor D. W.
A1 Leake, Mark C.
A1 Johnson, Steven
A1 Al-Soufi, Wajih
A1 Penedo, J. Carlos
A1 Quinn, Steven D.
A1 Novo, Mercedes
K1 Fluorescence imaging
K1 Single-molecule biophysics
AB 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
PB Nature Publishing Group
YR 2019
FD 2019
LK http://hdl.handle.net/10347/20224
UL http://hdl.handle.net/10347/20224
LA eng
NO Dalgarno, P.A., Juan-Colás, J., Hedley, G.J. et al. Unveiling the multi-step solubilization mechanism of sub-micron size vesicles by detergents. Sci Rep 9, 12897 (2019) doi:10.1038/s41598-019-49210-0
NO We thank Dr. Arvydas Ruseckas for helping with fluorescence lifetime measurements and the EPSRC (EP/P030017) for support
DS Minerva
RD 28 abr 2026