Sökning: onr:"swepub:oai:research.chalmers.se:74d884eb-182e-42c6-91a0-a52d936e71fe" > Preserved Transmemb...
Fältnamn | Indikatorer | Metadata |
---|---|---|
000 | 04032naa a2200397 4500 | |
001 | oai:research.chalmers.se:74d884eb-182e-42c6-91a0-a52d936e71fe | |
003 | SwePub | |
008 | 171008s2015 | |||||||||||000 ||eng| | |
024 | 7 | a https://doi.org/10.1021/acs.analchem.5b014492 DOI |
024 | 7 | a https://research.chalmers.se/publication/2234342 URI |
040 | a (SwePub)cth | |
041 | a engb eng | |
042 | 9 SwePub | |
072 | 7 | a art2 swepub-publicationtype |
072 | 7 | a ref2 swepub-contenttype |
100 | 1 | a Pace, Hudson,d 1982u Chalmers tekniska högskola,Chalmers University of Technology4 aut0 (Swepub:cth)hudsonp |
245 | 1 0 | a Preserved Transmembrane Protein Mobility in Polymer-Supported Lipid Bilayers Derived from Cell Membranes |
264 | c 2015-08-24 | |
264 | 1 | b American Chemical Society (ACS),c 2015 |
520 | a 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. | |
650 | 7 | a NATURVETENSKAPx Biologix Cellbiologi0 (SwePub)106042 hsv//swe |
650 | 7 | a NATURAL SCIENCESx Biological Sciencesx Cell Biology0 (SwePub)106042 hsv//eng |
650 | 7 | a NATURVETENSKAPx Biologix Biofysik0 (SwePub)106032 hsv//swe |
650 | 7 | a NATURAL SCIENCESx Biological Sciencesx Biophysics0 (SwePub)106032 hsv//eng |
700 | 1 | a Simonsson Nyström, Lisa,d 1982u Chalmers tekniska högskola,Chalmers University of Technology4 aut0 (Swepub:cth)slisa |
700 | 1 | a Gunnarsson, Anders,d 1981u AstraZeneca AB4 aut0 (Swepub:cth)kb01guan |
700 | 1 | a Eck, Elizabeth,d 1982u Chalmers tekniska högskola,Chalmers University of Technology4 aut0 (Swepub:cth)eeke |
700 | 1 | a Monson, C.u Southern Utah University4 aut |
700 | 1 | a Geschwindner, S.u AstraZeneca AB4 aut |
700 | 1 | a Snijder, Arjan,d 1971u AstraZeneca AB4 aut |
700 | 1 | a Höök, Fredrik,d 1966u Chalmers tekniska högskola,Chalmers University of Technology4 aut0 (Swepub:cth)fredrikh |
710 | 2 | a Chalmers tekniska högskolab AstraZeneca AB4 org |
773 | 0 | t Analytical Chemistryd : American Chemical Society (ACS)g 87:18, s. 9194-9203q 87:18<9194-9203x 0003-2700x 1520-6882 |
856 | 4 | u http://dx.doi.org/10.1021/acs.analchem.5b01449y FULLTEXT |
856 | 4 8 | u https://doi.org/10.1021/acs.analchem.5b01449 |
856 | 4 8 | u https://research.chalmers.se/publication/223434 |
Kungliga biblioteket hanterar dina personuppgifter i enlighet med EU:s dataskyddsförordning (2018), GDPR. Läs mer om hur det funkar här.
Så här hanterar KB dina uppgifter vid användning av denna tjänst.
Kopiera och spara länken för att återkomma till aktuell vy