Biomimetic membrane platform: Fabrication, characterization and applications
2013 (English)In: Colloids and Surfaces B: Biointerfaces, ISSN 0927-7765, E-ISSN 1873-4367, Vol. 103, 510-516 p.Article in journal (Refereed) Published
A facile method for assembly of biomimetic membranes serving as a platform for expression and insertion of membrane proteins is described. The membrane architecture was constructed in three steps: (i) assembly/printing of alpha-laminin peptide (P19) spacer on gold to separate solid support from the membrane architecture; (ii) covalent coupling of different lipid anchors to the P19 layer to serve as stabilizers of the inner leaflet during bilayer formation; (iii) lipid vesicle spreading to form a complete bilayer. Two different lipid membrane systems were examined and two different P19 architectures prepared by either self-assembly or mu-contact printing were tested and characterized using contact angle (CA) goniometry, surface plasmon resonance (SPR) spectroscopy and imaging surface plasmon resonance (iSPR). It is shown that surface coverage of cushion layer is significantly improved by mu-contact printing thereby facilitating bilayer formation as compared to self-assembly. To validate applicability of proposed methodology, incorporation of Cytochrome bo(3) ubiquinol oxidase (Cyt-bo(3)) into biomimetic membrane was performed by in vitro expression technique which was further monitored by surface plasmon enhanced fluorescence spectroscopy (SPFS). The results showed that solid supported planar membranes, tethered by alpha-laminin peptide cushion layer, provide an attractive environment for membrane protein insertion and characterization.
Place, publisher, year, edition, pages
Elsevier , 2013. Vol. 103, 510-516 p.
Tethered lipid membrane, Self-assembly, mu-Contact printing, alpha-Laminin peptide cushion, Cytochrome bo(3) ubiquinol oxidase
Medical and Health Sciences
IdentifiersURN: urn:nbn:se:liu:diva-90194DOI: 10.1016/j.colsurfb.2012.10.066ISI: 000315127000067OAI: oai:DiVA.org:liu-90194DiVA: diva2:612370
Funding Agencies|A*STAR JCOGrant of Singapore|10/03/FG/06/06|2013-03-212013-03-212013-03-21