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  Reconstitution of SNARE proteins into solid-supported lipid bilayer stacks and X-ray structure analysis.

Xu, Y., Kuhlmann, J., Brennich, M., Komorowski, K., Jahn, R., Steinem, C., et al. (2018). Reconstitution of SNARE proteins into solid-supported lipid bilayer stacks and X-ray structure analysis. Biochimica et Biophysica Acta (BBA) - Biomembranes, 1860(2), 566-578. doi:10.1016/j.bbamem.2017.10.023.

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 Urheber:
Xu, Y., Autor
Kuhlmann, J., Autor
Brennich, M., Autor
Komorowski, K., Autor
Jahn, R.1, Autor           
Steinem, C., Autor
Salditt, T., Autor
Affiliations:
1Department of Neurobiology, MPI for biophysical chemistry, Max Planck Society, ou_578595              

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Schlagwörter: SNARE reconstitution; Supported lipid bilayer stack; Micelle and vesicle; SAXS; X-ray reflectivity; GISAXS
 Zusammenfassung: SNAREs are known as an important family of proteins mediating vesicle fusion. For various biophysical studies, they have been reconstituted into supported single bilayers via proteoliposome adsorption and rupture. In this study we extended this method to the reconstitution of SNAREs into supported multilamellar lipid membranes, i.e. oriented multibilayer stacks, as an ideal model system for X-ray structure analysis (X-ray reflectivity and diffraction). The reconstitution was implemented through a pathway of proteomicelle, proteoliposome and multibilayer. To monitor the structural evolution in each step, we used small-angle X-ray scattering for the proteomicelles and proteoliposomes, followed by X-ray reflectivity and grazing-incidence small-angle scattering for the multibilayers. Results show that SNAREs can be successfully reconstituted into supported multibilayers, with high enough orientational alignment for the application of surface sensitive X-ray characterizations. Based on this protocol, we then investigated the effect of SNAREs on the structure and phase diagram of the lipid membranes. Beyond this application, this reconstitution protocol could also be useful for X-ray analysis of many further membrane proteins.

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Sprache(n): eng - English
 Datum: 2017-11-282018-02
 Publikationsstatus: Erschienen
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 Ort, Verlag, Ausgabe: -
 Inhaltsverzeichnis: -
 Art der Begutachtung: Expertenbegutachtung
 Identifikatoren: DOI: 10.1016/j.bbamem.2017.10.023
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Titel: Biochimica et Biophysica Acta (BBA) - Biomembranes
Genre der Quelle: Zeitschrift
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Ort, Verlag, Ausgabe: -
Seiten: - Band / Heft: 1860 (2) Artikelnummer: - Start- / Endseite: 566 - 578 Identifikator: -