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

Yue, Ke ; Trung, Tran Nam ; Zhu, Yiyong ; Kaldenhoff, Ralf ; Kai, Lei (2023)
Co-Translational Insertion of Aquaporins into Liposome for Functional Analysis via an E. coli Based Cell-Free Protein Synthesis System.
In: Cells, 2019, 8 (11)
doi: 10.26083/tuprints-00015747
Artikel, Zweitveröffentlichung, Verlagsversion

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Kurzbeschreibung (Abstract)

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.

Typ des Eintrags: Artikel
Erschienen: 2023
Autor(en): Yue, Ke ; Trung, Tran Nam ; Zhu, Yiyong ; Kaldenhoff, Ralf ; Kai, Lei
Art des Eintrags: Zweitveröffentlichung
Titel: Co-Translational Insertion of Aquaporins into Liposome for Functional Analysis via an E. coli Based Cell-Free Protein Synthesis System
Sprache: Englisch
Publikationsjahr: 1 Dezember 2023
Ort: Darmstadt
Publikationsdatum der Erstveröffentlichung: 2019
Ort der Erstveröffentlichung: Basel
Verlag: MDPI
Titel der Zeitschrift, Zeitung oder Schriftenreihe: Cells
Jahrgang/Volume einer Zeitschrift: 8
(Heft-)Nummer: 11
Kollation: 11 Seiten
DOI: 10.26083/tuprints-00015747
URL / URN: https://tuprints.ulb.tu-darmstadt.de/15747
Zugehörige Links:
Herkunft: Zweitveröffentlichung DeepGreen
Kurzbeschreibung (Abstract):

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.

Freie Schlagworte: aquaporin, cell-free protein synthesis, co-translational insertion, proteo-liposome
Status: Verlagsversion
URN: urn:nbn:de:tuda-tuprints-157472
Zusätzliche Informationen:

This article belongs to the Special Issue Aquaporins 2019

Sachgruppe der Dewey Dezimalklassifikatin (DDC): 500 Naturwissenschaften und Mathematik > 570 Biowissenschaften, Biologie
Fachbereich(e)/-gebiet(e): 10 Fachbereich Biologie
10 Fachbereich Biologie > Applied Plant Sciences
Hinterlegungsdatum: 01 Dez 2023 14:07
Letzte Änderung: 04 Dez 2023 12:22
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