Baumann, A; Kerruth, S; Fitter, J; Büldt, G; Heberle, J; Schlesinger, R; Ataka, K
(2016)
In-Situ Observation of Membrane Protein Folding during Cell-Free Expression.
PLoS One, 11 (3).
e0151051.
ISSN 1932-6203
https://doi.org/10.1371/journal.pone.0151051
SGUL Authors: Kerruth, Silke
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Abstract
Proper insertion, folding and assembly of functional proteins in biological membranes are key processes to warrant activity of a living cell. Here, we present a novel approach to trace folding and insertion of a nascent membrane protein leaving the ribosome and penetrating the bilayer. Surface Enhanced IR Absorption Spectroscopy selectively monitored insertion and folding of membrane proteins during cell-free expression in a label-free and non-invasive manner. Protein synthesis was performed in an optical cell containing a prism covered with a thin gold film with nanodiscs on top, providing an artificial lipid bilayer for folding. In a pilot experiment, the folding pathway of bacteriorhodopsin via various secondary and tertiary structures was visualized. Thus, a methodology is established with which the folding reaction of other more complex membrane proteins can be observed during protein biosynthesis (in situ and in operando) at molecular resolution.
Item Type: | Article | ||||||
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Additional Information: | © 2016 Baumann et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. | ||||||
Keywords: | Cell-Free System, Membrane Proteins, Protein Folding, Spectrophotometry, Ultraviolet, Cell-Free System, Membrane Proteins, Spectrophotometry, Ultraviolet, Protein Folding, General Science & Technology, MD Multidisciplinary | ||||||
SGUL Research Institute / Research Centre: | Academic Structure > Molecular and Clinical Sciences Research Institute (MCS) | ||||||
Journal or Publication Title: | PLoS One | ||||||
ISSN: | 1932-6203 | ||||||
Language: | eng | ||||||
Dates: |
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Publisher License: | Creative Commons: Attribution 4.0 | ||||||
PubMed ID: | 26978519 | ||||||
Go to PubMed abstract | |||||||
URI: | https://openaccess.sgul.ac.uk/id/eprint/109128 | ||||||
Publisher's version: | https://doi.org/10.1371/journal.pone.0151051 |
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