English
 
Help Privacy Policy Disclaimer
  Advanced SearchBrowse

Item

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

Yue, K., Tran Trung, N., Zhu, Y., Kaldenhoff, R., & Kai, L. (2019). Co-Translational Insertion of Aquaporins into Liposome for Functional Analysis via an E. coli Based Cell-Free Protein Synthesis System. CELLS, 8(11): 1325. doi:10.3390/cells8111325.

Item is

Basic

show hide
Genre: Journal Article

Files

show Files
hide Files
:
cells-08-01325.pdf (Any fulltext), 2MB
Name:
cells-08-01325.pdf
Description:
-
Visibility:
Public
MIME-Type / Checksum:
application/pdf / [MD5]
Technical Metadata:
Copyright Date:
-
Copyright Info:
open access article
License:
-

Locators

show

Creators

show
hide
 Creators:
Yue, Ke1, Author
Tran Trung, Nam1, Author
Zhu, Yiyong1, Author
Kaldenhoff, Ralf1, Author
Kai, Lei2, Author              
Affiliations:
1external, ou_persistent22              
2Schwille, Petra / Cellular and Molecular Biophysics, Max Planck Institute of Biochemistry, Max Planck Society, ou_1565169              

Content

show
hide
Free keywords: INTEGRAL MEMBRANE-PROTEINS; FREE EXPRESSION; WATER CHANNELS; RECONSTITUTION; STABILIZATION; DETERGENTS; MECHANISM; GLYCEROL; AQPZaquaporin; cell-free protein synthesis; co-translational insertion; proteo-liposome;
 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.

Details

show
hide
Language(s): eng - English
 Dates: 2019
 Publication Status: Published online
 Pages: 11
 Publishing info: -
 Table of Contents: -
 Rev. Type: Peer
 Identifiers: ISI: 000502266700023
DOI: 10.3390/cells8111325
 Degree: -

Event

show

Legal Case

show

Project information

show

Source 1

show
hide
Title: CELLS
Source Genre: Journal
 Creator(s):
Affiliations:
Publ. Info: ST ALBAN-ANLAGE 66, CH-4052 BASEL, SWITZERLAND : MDPI
Pages: - Volume / Issue: 8 (11) Sequence Number: 1325 Start / End Page: - Identifier: -