Search: id:"swepub:oai:research.chalmers.se:74d884eb-182e-42c6-91a0-a52d936e71fe" >
Preserved Transmemb...
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Pace, Hudson,1982Chalmers tekniska högskola,Chalmers University of Technology
(author)
Preserved Transmembrane Protein Mobility in Polymer-Supported Lipid Bilayers Derived from Cell Membranes
- Article/chapterEnglish2015
Publisher, publication year, extent ...
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2015-08-24
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American Chemical Society (ACS),2015
Numbers
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LIBRIS-ID:oai:research.chalmers.se:74d884eb-182e-42c6-91a0-a52d936e71fe
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https://doi.org/10.1021/acs.analchem.5b01449DOI
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https://research.chalmers.se/publication/223434URI
Supplementary language notes
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Language:English
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Summary in:English
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Subject category:art swepub-publicationtype
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Subject category:ref swepub-contenttype
Notes
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Supported lipid bilayers (SLBs) have contributed invaluable information about the physiochemical properties of cell membranes, but their compositional simplicity often limits the level of knowledge that can be gained about the structure and function of transmembrane proteins in their native environment. Herein, we demonstrate a generic protocol for producing polymer-supported lipid bilayers on glass surfaces that contain essentially all naturally occurring cell-membrane components of a cell line while still retaining transmembrane protein mobility and activity. This was achieved by merging vesicles made from synthetic lipids (PEGylated lipids and POPC lipids) with native cell-membrane vesicles to generate hybrid vesicles which readily rupture into a continuous polymer-supported lipid bilayer. To investigate the properties of these complex hybrid SLBs and particularly the behavior of their integral membrane-proteins, we used total internal reflection fluorescence imaging to study a transmembrane protease, β-secretase 1 (BACE1), whose ectoplasmic and cytoplasmic domains could both be specifically targeted with fluorescent reporters. By selectively probing the two different orientations of BACE1 in the resulting hybrid SLBs, the role of the PEG-cushion on transmembrane protein lateral mobility was investigated. The results reveal the necessity of having the PEGylated lipids present during vesicle adsorption to prevent immobilization of transmembrane proteins with protruding domains. The proteolytic activity of BACE1 was unadulterated by the sonication process used to merge the synthetic and native membrane vesicles; importantly it was also conserved in the SLB. The presented strategy could thus serve both fundamental studies of membrane biophysics and the production of surface-based bioanalytical sensor platforms.
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Simonsson Nyström, Lisa,1982Chalmers tekniska högskola,Chalmers University of Technology(Swepub:cth)slisa
(author)
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Gunnarsson, Anders,1981AstraZeneca AB(Swepub:cth)kb01guan
(author)
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Eck, Elizabeth,1982Chalmers tekniska högskola,Chalmers University of Technology(Swepub:cth)eeke
(author)
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Monson, C.Southern Utah University
(author)
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Geschwindner, S.AstraZeneca AB
(author)
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Snijder, Arjan,1971AstraZeneca AB
(author)
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Höök, Fredrik,1966Chalmers tekniska högskola,Chalmers University of Technology(Swepub:cth)fredrikh
(author)
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Chalmers tekniska högskolaAstraZeneca AB
(creator_code:org_t)
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In:Analytical Chemistry: American Chemical Society (ACS)87:18, s. 9194-92030003-27001520-6882
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Monson, C.
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Geschwindner, S.
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NATURAL SCIENCES
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and Cell Biology
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and Biophysics
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Analytical Chemi ...
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Chalmers University of Technology