AUTHOR=Blackholly Laura R. , Harris Nicola J. , Findlay Heather E. , Booth Paula J. TITLE=Cell-Free Expression to Probe Co-Translational Insertion of an Alpha Helical Membrane Protein JOURNAL=Frontiers in Molecular Biosciences VOLUME=Volume 9 - 2022 YEAR=2022 URL=https://www.frontiersin.org/journals/molecular-biosciences/articles/10.3389/fmolb.2022.795212 DOI=10.3389/fmolb.2022.795212 ISSN=2296-889X ABSTRACT=
The majority of alpha helical membrane proteins fold co-translationally during their synthesis on the ribosome. In contrast, most mechanistic folding studies address refolding of full-length proteins from artificially induced denatured states that are far removed from the natural co-translational process. Cell-free translation of membrane proteins is emerging as a useful tool to address folding during translation by a ribosome. We summarise the benefits of this approach and show how it can be successfully extended to a membrane protein with a complex topology. The bacterial leucine transporter, LeuT can be synthesised and inserted into lipid membranes using a variety of