date: 2019-10-27T09:40:33Z
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pdf:docinfo:title: Co-Translational Insertion of Aquaporins into Liposome for Functional Analysis via an E. coli Based Cell-Free Protein Synthesis System
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subject: 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.
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dc:title: Co-Translational Insertion of Aquaporins into Liposome for Functional Analysis via an E. coli Based Cell-Free Protein Synthesis System
modified: 2019-10-27T09:40:33Z
cp:subject: 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.
pdf:docinfo:subject: 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.
pdf:docinfo:creator: Ke Yue, Tran Nam Trung, Yiyong Zhu, Ralf Kaldenhoff and Lei Kai
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meta:author: Ke Yue, Tran Nam Trung, Yiyong Zhu, Ralf Kaldenhoff and Lei Kai
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Creation-Date: 2019-10-27T09:40:33Z
Author: Ke Yue, Tran Nam Trung, Yiyong Zhu, Ralf Kaldenhoff and Lei Kai
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Keywords: aquaporin; cell-free protein synthesis; co-translational insertion; proteo-liposome
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dc:creator: Ke Yue, Tran Nam Trung, Yiyong Zhu, Ralf Kaldenhoff and Lei Kai
dcterms:created: 2019-10-27T09:40:33Z
Last-Modified: 2019-10-27T09:40:33Z
dcterms:modified: 2019-10-27T09:40:33Z
title: Co-Translational Insertion of Aquaporins into Liposome for Functional Analysis via an E. coli Based Cell-Free Protein Synthesis System
Last-Save-Date: 2019-10-27T09:40:33Z
pdf:docinfo:keywords: aquaporin; cell-free protein synthesis; co-translational insertion; proteo-liposome
pdf:docinfo:modified: 2019-10-27T09:40:33Z
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creator: Ke Yue, Tran Nam Trung, Yiyong Zhu, Ralf Kaldenhoff and Lei Kai
dc:subject: aquaporin; cell-free protein synthesis; co-translational insertion; proteo-liposome
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meta:keyword: aquaporin; cell-free protein synthesis; co-translational insertion; proteo-liposome
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pdf:docinfo:created: 2019-10-27T09:40:33Z