Berlin 2012 – wissenschaftliches Programm
Bereiche | Tage | Auswahl | Suche | Aktualisierungen | Downloads | Hilfe
BP: Fachverband Biologische Physik
BP 26: Posters: Membranes and Vesicles
BP 26.10: Poster
Donnerstag, 29. März 2012, 17:30–19:30, Poster A
How to tailor giant proteoliposomes — •Susanne F. Fenz1, Rita Sachse2, Stefan Kubick2, and Thomas Schmidt1 — 1LION, Leiden University, The Netherlands — 2Zellulaere Biotechnologie, Fraunhofer IBMT, Potsdam-Golm, Germany
In this project we address the challenge to incorporate transmembrane proteins in the membrane of giant unilamellar vesicles (GUVs). The reconstitution of biologically relevant transmembrane proteins, like receptors or channel proteins, into GUVs makes them easily accessible to further biochemical and physical investigation. Our strategy combines two approaches: in vitro eukaryotic protein expression and electro-swelling. The in vitro protein expression system is based on insect lysates. It provides endoplasmic reticulum (ER)-based vesicles which enable signal-induced translocation and post-translational modification. Starting from these vesicles of approximately 1 mum diameter we applied electro-swelling to achieve giant proteoliposomes. Our recent work showed that the efficiency of this method can be improved substantially by the presence of synthetic lipids in the electro-swelling process. As an example, we introduced the one-transmembrane protein heparin-binding epidermal growth factor-like factor Hb-EGF-eYFP in GUV membranes aided by the lipid DOPC. We applied single-molecule fluorescence microscopy to detect and further characterize the protein. In addition, we introduced biotinylated lipids that enabled us to immobilize the protein-decorated GUVs to streptavidin coated surfaces. We envision this achievement as an important first step toward systematic protein studies on technical surfaces.