University of Natural Resources and Life Sciences, Vienna (BOKU) - Research portal

Logo BOKU Resarch Portal

Selected Publication:

Yildiz, AA; Knoll, W; Gennis, RB; Sinner, EK.
(2012): Cell-free synthesis of cytochrome bo(3) ubiquinol oxidase in artificial membranes.
Anal Biochem. 2012; 423(1):39-45 FullText FullText_BOKU

Abstract:
The analysis of membrane proteins is notoriously difficult because isolation and detergent-mediated reconstitution often results in compromising the protein structure and function. We introduce a novel strategy of combining a cell-free expression method for synthesis of a protein species coping with one of the most important obstacles in membrane protein research preserving the structural-functional integrity of a membrane protein species and providing a stable matrix for application of analytical tools to characterize the membrane protein of interest. We address integration and subsequent characterization of the cytochrome bo(3) ubiquinol oxidase (Cyt-bo(3)) from de novo synthesis without the effort of conventional cell culture, isolation, and purification procedures. The experimental output supports our idea of a suitable platform for in vitro protein synthesis and functional integration into a membrane-mimicking structure. We show the compatibility of different concepts of in vitro synthesis toward biosensor applicability by the example of Cyt-bo(3) protein expression. Our results obtained from in vitro synthesized proteins displayed similar behavior to proteins isolated from the cellular context. Overall, our approach is suitable for the in vitro expression of "complex" protein species such as Cyt-bo(3), which can be reproducible and stably synthesized and preserved in robust, synthetic planar membrane architecture. (C) 2012 Elsevier Inc. All rights reserved.
Authors BOKU Wien:
Ehmoser Eva-Kathrin

Find related publications in this database (Keywords)
Cell-free protein synthesis
Cytochrome bo(3) ubiquinol oxidase
Electron transport chain
Mitochondrial disorders
Artificial membrane
Synthetic bioarchitectures


Altmetric:
© BOKU Wien Imprint