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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 ; Tran, Nam Trung ; Zhu, Yiyong ; Kaldenhoff, Ralf ; Kai, Lei (2019)
Co-Translational Insertion of Aquaporins into Liposome for Functional Analysis via an E. coli Based Cell-Free Protein Synthesis System.
In: Cells, 8 (11)
doi: 10.3390/cells8111325
Artikel, Bibliographie

Dies ist die neueste Version dieses Eintrags.

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 and 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 PIP2;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: 2019
Autor(en): Yue, Ke ; Tran, Nam Trung ; Zhu, Yiyong ; Kaldenhoff, Ralf ; Kai, Lei
Art des Eintrags: Bibliographie
Titel: Co-Translational Insertion of Aquaporins into Liposome for Functional Analysis via an E. coli Based Cell-Free Protein Synthesis System.
Sprache: Englisch
Publikationsjahr: 27 Oktober 2019
Titel der Zeitschrift, Zeitung oder Schriftenreihe: Cells
Jahrgang/Volume einer Zeitschrift: 8
(Heft-)Nummer: 11
DOI: 10.3390/cells8111325
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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 and 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 PIP2;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.

ID-Nummer: pmid:31717877
Fachbereich(e)/-gebiet(e): 10 Fachbereich Biologie
10 Fachbereich Biologie > Applied Plant Sciences
Hinterlegungsdatum: 18 Nov 2019 11:42
Letzte Änderung: 04 Dez 2023 12:23
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