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

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  • معلومة اضافية
    • Contributors:
      Universidade de Santiago de Compostela. Departamento de Química Física
    • بيانات النشر:
      Nature Publishing Group
    • Collection:
      Minerva - Repositorio institucional da Universidade de Santiago de Compostela (USC)
    • نبذة مختصرة :
      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 ; We thank Dr. Arvydas Ruseckas for helping with fluorescence lifetime measurements and the EPSRC (EP/P030017) for support ; SI
    • File Description:
      application/pdf
    • ISSN:
      2045-2322
    • Relation:
      https://doi.org/10.1038/s41598-019-49210-0; 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; http://hdl.handle.net/10347/20224
    • الرقم المعرف:
      10.1038/s41598-019-49210-0
    • الدخول الالكتروني :
      http://hdl.handle.net/10347/20224
      https://doi.org/10.1038/s41598-019-49210-0
    • Rights:
      © The Author(s) 2019. Open Access. This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. Te images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0 ; Atribución 4.0 Internacional ; http://creativecommons.org/licenses/by/4.0/ ; info:eu-repo/semantics/openAccess
    • الرقم المعرف:
      edsbas.1CF8D091