In Situ Monitoring of Membrane Protein Insertion into Block Copolymer Vesicle Membranes and Their Spreading via Potential-Assisted Approach
- Tayebeh Mirzaei GarakaniTayebeh Mirzaei GarakaniInstitute of Biotechnology, RWTH Aachen University, Worringer Weg 3, D-52074 Aachen, GermanyDWI - Leibniz Institute for Interactive Materials, Forckenbeckstraße 50, D-52074, Aachen, GermanyMore by Tayebeh Mirzaei Garakani
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- Zhanzhi LiuZhanzhi LiuInstitute of Biotechnology, RWTH Aachen University, Worringer Weg 3, D-52074 Aachen, GermanyMore by Zhanzhi Liu
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- Ulrich GlebeUlrich GlebeFraunhofer Institute for Applied Polymer Research IAP, Geiselbergstraße 69, 14476 Potsdam-Golm, GermanyChair of Polymer Materials and Polymer Technologies, Institute of Chemistry, University of Potsdam, Karl-Liebknecht-Str. 24-25, 14476 Potsdam-Golm, GermanyMore by Ulrich Glebe
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- Julia GehrmannJulia GehrmannInstitute of Biotechnology, RWTH Aachen University, Worringer Weg 3, D-52074 Aachen, GermanyMore by Julia Gehrmann
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- Jaroslav LazarJaroslav LazarInstitute of Materials in Electrical Engineering 1, RWTH Aachen University, Sommerfeldstraße 24, 52074 Aachen, GermanyMore by Jaroslav Lazar
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- Marie Anna Stephanie MertensMarie Anna Stephanie MertensInstitute of Biotechnology, RWTH Aachen University, Worringer Weg 3, D-52074 Aachen, GermanyMore by Marie Anna Stephanie Mertens
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- Mieke MöllerMieke MöllerInstitute of Biotechnology, RWTH Aachen University, Worringer Weg 3, D-52074 Aachen, GermanyMore by Mieke Möller
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- Niloofar HamzeluiNiloofar HamzeluiInstitute of Biotechnology, RWTH Aachen University, Worringer Weg 3, D-52074 Aachen, GermanyMore by Niloofar Hamzelui
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- Leilei ZhuLeilei ZhuInstitute of Biotechnology, RWTH Aachen University, Worringer Weg 3, D-52074 Aachen, GermanyMore by Leilei Zhu
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- Uwe SchnakenbergUwe SchnakenbergInstitute of Materials in Electrical Engineering 1, RWTH Aachen University, Sommerfeldstraße 24, 52074 Aachen, GermanyMore by Uwe Schnakenberg
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- Alexander BökerAlexander BökerFraunhofer Institute for Applied Polymer Research IAP, Geiselbergstraße 69, 14476 Potsdam-Golm, GermanyChair of Polymer Materials and Polymer Technologies, Institute of Chemistry, University of Potsdam, Karl-Liebknecht-Str. 24-25, 14476 Potsdam-Golm, GermanyMore by Alexander Böker
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- Ulrich Schwaneberg*Ulrich Schwaneberg*E-mail: [email protected]Institute of Biotechnology, RWTH Aachen University, Worringer Weg 3, D-52074 Aachen, GermanyDWI - Leibniz Institute for Interactive Materials, Forckenbeckstraße 50, D-52074, Aachen, GermanyMore by Ulrich Schwaneberg
Abstract
Synthosomes are polymer vesicles with transmembrane proteins incorporated into block copolymer membranes. They have been used for selective transport in or out of the vesicles as well as catalysis inside the compartments. However, both the insertion process of the membrane protein, forming nanopores, and the spreading of the vesicles on planar substrates to form solid-supported biomimetic membranes have been rarely studied yet. Herein, we address these two points and, first, shed light on the real-time monitoring of protein insertion via isothermal titration calorimetry. Second, the spreading process on different solid supports, namely, SiO2, glass, and gold, via different techniques like spin- and dip-coating as well as a completely new approach of potential-assisted spreading on gold surfaces was studied. While inhomogeneous layers occur via traditional methods, our proposed potential-assisted strategy to induce adsorption of positively charged vesicles by applying negative potential on the electrode leads to remarkable vesicle spreading and their further fusion to form more homogeneous planar copolymer films on gold. The polymer vesicles in our study are formed from amphiphilic copolymers poly(2-methyl oxazoline)-block-poly(dimethylsiloxane)-block-poly(2-methyl oxazoline) (PMOXA-b-PDMS-b-PMOXA). Engineered variants of the transmembrane protein ferric hydroxamate uptake protein component A (FhuA), one of the largest β-barrel channel proteins, are used as model nanopores. The incorporation of FhuA Δ1-160 is shown to facilitate the vesicle spreading process further. Moreover, high accessibility of cysteine inside the channel was proven by linkage of a fluorescent dye inside the engineered variant FhuA ΔCVFtev and hence preserved functionality of the channels after spreading. The porosity and functionality of the spread synthosomes on the gold plates have been examined by studying the passive ion transport response in the presence of Li+ and ClO4– ions and electrochemical impedance spectroscopy analysis. Our approach to form solid-supported biomimetic membranes via the potential-assisted strategy could be important for the development of new (bio-) sensors and membranes.
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This article is cited by 12 publications.
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