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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