Your browser doesn't support javascript.
loading
Unveiling the multi-step solubilization mechanism of sub-micron size vesicles by detergents.
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.
Afiliación
  • Dalgarno PA; SUPA School of Physics and Astronomy, University of St. Andrews, North Haugh, Fife, KY16 9SS, UK.
  • Juan-Colás J; Institute of Biological Physics and Bioengineering, School of Engineering and Physical Sciences, Heriot-Watt University, Edinburgh, EH14 4AS, UK.
  • Hedley GJ; Department of Electronic Engineering, University of York, Heslington, York, YO10 5DD, UK.
  • Piñeiro L; SUPA School of Physics and Astronomy, University of St. Andrews, North Haugh, Fife, KY16 9SS, UK.
  • Novo M; School of Chemistry, University of Glasgow, Glasgow, Scotland, G12 8QQ, United Kingdom.
  • Perez-Gonzalez C; Department of Physical Chemistry, Faculty of Science, University of Santiago de Compostela, Lugo, E-27002, Spain.
  • Samuel IDW; Department of Physical Chemistry, Faculty of Science, University of Santiago de Compostela, Lugo, E-27002, Spain.
  • Leake MC; SUPA School of Physics and Astronomy, University of St. Andrews, North Haugh, Fife, KY16 9SS, UK.
  • Johnson S; SUPA School of Physics and Astronomy, University of St. Andrews, North Haugh, Fife, KY16 9SS, UK.
  • Al-Soufi W; Department of Physics, University of York, Heslington, York, England, YO10 5DD, UK.
  • Penedo JC; Department of Biology, University of York, Heslington, York, YO10 5DD, UK.
  • Quinn SD; Department of Electronic Engineering, University of York, Heslington, York, YO10 5DD, UK.
Sci Rep ; 9(1): 12897, 2019 09 09.
Article en En | MEDLINE | ID: mdl-31501469
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.

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Idioma: En Revista: Sci Rep Año: 2019 Tipo del documento: Article Pais de publicación: Reino Unido

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Idioma: En Revista: Sci Rep Año: 2019 Tipo del documento: Article Pais de publicación: Reino Unido