Cell-free expression tools to study co-translational folding of alpha helical membrane transporters.

ABSTRACT: Most helical membrane proteins fold co-translationally during unidirectional polypeptide elongation by the ribosome. Studies thus far, however, have largely focussed on refolding full-length proteins from artificially induced denatured states that are far removed from the natural co-translational process. Cell-free translation offers opportunities to remedy this deficit in folding studies and has previously been used for membrane proteins. We exploit this cell-free approach to develop tools to probe co-translational folding. We show that two transporters from the ubiquitous Major Facilitator Superfamily can successfully insert into a synthetic bilayer without the need for translocon insertase apparatus that is essential in vivo. We also assess the cooperativity of domain insertion, by expr

SUBMITTER: Harris NJ 

PROVIDER: S-EPMC7272624 | biostudies-literature | 2020 Jun

REPOSITORIES: biostudies-literature