Abstract
Proteo-lipobeads (PLBs) are investigated as cell-free model systems to encapsulate membrane proteins
such as ion channels and transporters. PLBs are based on nickel nitrile tri-acetic acid (Ni-NTA)-
functionalized agarose beads, onto which membrane proteins (MP) are bound via histidine(his)-tag.
Composite beads thus obtained (subsequently called proteobeads) are dialyzed in the presence of lipid
micelles to form PLBs. As an example we employed cytochrome c oxidase from P. denitrificans with a
his-tag fused to the C-terminus of subunit I. In this orientation the P side of CcO faces the outside of
the PLB and hence protons are released to the outer aqueous phase, when electron transfer is initiated
by light excitation of Ru complexes. Proton release kinetics was probed by fluorescence microscopy using
the pH-sensitive sensor molecule fluorescein DHPE inserted into the lipid layer. In order to monitor the
generation of membrane potentials we performed a FLIPR assay on the CcO embedded in PLBs using the
FRET pair CC2-DMPE/DiSBAC2(3). The combined results show that PLBs can be used as a model system
Originalsprache | Englisch |
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Seiten (von - bis) | 119-125 |
Seitenumfang | 7 |
Fachzeitschrift | Journal of Colloid & Interface Science |
Volume | 500 |
DOIs | |
Publikationsstatus | Veröffentlicht - 2017 |
Research Field
- Biosensor Technologies