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Prokaryotic Aquaporins
Open AccessArticle

Co-Translational Insertion of Aquaporins into Liposome for Functional Analysis via an E. coli Based Cell-Free Protein Synthesis System

1
The Key Laboratory of Biotechnology for Medicinal Plants of Jiangsu Province, School of Life Sciences, Jiangsu Normal University, Xuzhou 22116, China
2
Department of Biology, Applied Plant Sciences, Technische Universität Darmstadt, Schnittspahn Strasse 10, D-64287 Darmstadt, Germany
3
Jiangsu Provincial Key Lab for Organic Solid Waste Utilization, National Engineering Research Center for Organic-based Fertilizers, Jiangsu Collaborative Innovation Center for Solid Organic Waste Resource Utilization, Nanjing Agricultural University, Nanjing 210095, China
4
Department of Cellular and Molecular Biophysics, Max Planck Institute of Biochemistry, D-82152 Martinsried, Germany
*
Author to whom correspondence should be addressed.
Cells 2019, 8(11), 1325; https://doi.org/10.3390/cells8111325
Received: 30 September 2019 / Revised: 24 October 2019 / Accepted: 24 October 2019 / Published: 27 October 2019
(This article belongs to the Special Issue Aquaporins 2019)
Aquaporins are important and well-studied water channel membrane proteins. However, being membrane proteins, sample preparation for functional analysis is tedious and time-consuming. In this paper, we report a new approach for the co-translational insertion of two aquaporins from Escherichia coli and Nicotiana tabacum using the CFPS system. This was done in the presence of liposomes with a modified procedure to form homogenous proteo-liposomes suitable for functional analysis of water permeability using stopped-flow spectrophotometry. Two model aquaporins, AqpZ and NtPIP2;1, were successfully incorporated into the liposome in their active forms. Shifted green fluorescent protein was fused to the C-terminal part of AqpZ to monitor its insertion and status in the lipid environment. This new fast approach offers a fast and straightforward method for the functional analysis of aquaporins in both prokaryotic and eukaryotic organisms. View Full-Text
Keywords: aquaporin; cell-free protein synthesis; co-translational insertion; proteo-liposome aquaporin; cell-free protein synthesis; co-translational insertion; proteo-liposome
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MDPI and ACS Style

Yue, K.; Trung, T.N.; Zhu, Y.; Kaldenhoff, R.; Kai, L. Co-Translational Insertion of Aquaporins into Liposome for Functional Analysis via an E. coli Based Cell-Free Protein Synthesis System. Cells 2019, 8, 1325.

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