Regensburg 2004 – scientific programme
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AKB: Biologische Physik
AKB 50: Poster Session "Biological Physics"
AKB 50.122: Poster
Friday, March 12, 2004, 10:30–13:00, B
Deposition of Supported Membranes with F1F0-ATP-Synthase on Cellulose Thin Films — •Murat Tutus1, Thomas Nawroth2, and Motomu Tanaka1 — 1Biophysics Lab, Tech. Univ. Munich — 2Dept. Phys. E17, Tech. Univ. Munich
A new method for the reconstitution of proteins (F1F0-ATP-Synthase of Micrococcus luteus) into lipid vesicles without causing membrane disruption is developed. To verify non-disruptive, orientation selective reconstitution of ATP-Synthase in lipid vesicles, the vesicle size before and after insertion is quantitatively compared by dynamic light scattering experiments. To determine the side selectivity the proteoliposomes were spread onto biocompatible ultra thin (thickness of 5 to 10 nm) polymer supports where the homogeneity and the lateral mobility are characterized with fluorescence microscopy and fluorescence recovery after photobleaching. The protein distribution is checked by covalent coupling of synthetic dyes as well as by immunofluorescence labeling with antibodies. The glass slides coated with ultra thin cellulose films serve as interlayer to achieve homogeneous distribution over macroscopically large (cm2 order) surfaces. Moreover, immuno-labeling of F1 head group verifies the reconstitution of the entire protein into the supported membrane.