biostudies-literatureIkujuni APNational Institute of General Medical SciencesAmerican-Scandinavian FoundationNIGMS NIH HHS1185-1197https://www.ebi.ac.uk/biostudies/studies/S-EPMC10111266biostudies-literatureUnknown122(7)TolC is the trimeric outer membrane component of the efflux pump system in Escherichia coli that is responsible for antibiotic efflux from bacterial cells. Overexpression of efflux pumps has been reported to decrease susceptibility to antibiotics in a variety of bacterial pathogens. Reliable production of membrane proteins allows for the biophysical and structural characterization needed to better understand efflux and for the development of therapeutics. Preparation of recombinant protein for biochemical/structural studies often involves the production of proteins as inclusion body aggregates from which active proteins are recovered. Here, we find that the in vitro folding of TolC into its functional trimeric state from inclusion bodies is dependent on the headgroup composition of detergeBiophysical journalDetergent headgroups control TolC folding in vitro.PMC10111266P20 GM113117P20 GM103418DP2GM128201DP2 GM128201P20GM113117P20GM103418K12 GM0636512K12GM063651Slusky JSGDhar RIkujuni APBudiardjo SJfalseDetergent headgroups control TolC folding in vitro.TolC is the trimeric outer membrane component of the efflux pump system in Escherichia coli that is responsible for antibiotic efflux from bacterial cells. Overexpression of efflux pumps has been reported to decrease susceptibility to antibiotics in a variety of bacterial pathogens. Reliable production of membrane proteins allows for the biophysical and structural characterization needed to better understand efflux and for the development of therapeutics. Preparation of recombinant protein for biochemical/structural studies often involves the production of proteins as inclusion body aggregates from which active proteins are recovered. Here, we find that the in vitro folding of TolC into its functional trimeric state from inclusion bodies is dependent on the headgroup composition of deterge2023-01-01T00:00:00Z2023 Apr2026-05-26T14:05:16.797Z2025-02-18T22:34:35.054ZS-EPMC101112663677279610.1016/j.bpj.2023.02.007