Dataset Information

Molecular basis for the folding of ?-helical autotransporter passenger domains.

ABSTRACT: Bacterial autotransporters comprise a C-terminal ?-barrel domain, which must be correctly folded and inserted into the outer membrane to facilitate translocation of the N-terminal passenger domain to the cell exterior. Once at the surface, the passenger domains of most autotransporters are folded into an elongated ?-helix. In a cellular context, key molecules catalyze the assembly of the autotransporter ?-barrel domain. However, how the passenger domain folds into its functional form is poorly understood. Here we use mutational analysis on the autotransporter Pet to show that the ?-hairpin structure of the fifth extracellular loop of the ?-barrel domain has a crucial role for passenger domain folding into a ?-helix. Bioinformatics and structural analyses, and mutagenesis of a homologous autotransporter, suggest that this function is conserved among autotransporter proteins with ?-helical passenger domains. We propose that the autotransporter ?-barrel domain is a folding vector that nucleates folding of the passenger domain.

SUBMITTER: Yuan X

PROVIDER: S-EPMC5895577 | biostudies-literature | 2018 Apr

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

Molecular basis for the folding of β-helical autotransporter passenger domains.

Yuan Xiaojun X Johnson Matthew D MD Zhang Jing J Lo Alvin W AW Schembri Mark A MA Wijeyewickrema Lakshmi C LC Pike Robert N RN Huysmans Gerard H M GHM Henderson Ian R IR Leyton Denisse L DL

Nature communications 20180411 1

Bacterial autotransporters comprise a C-terminal β-barrel domain, which must be correctly folded and inserted into the outer membrane to facilitate translocation of the N-terminal passenger domain to the cell exterior. Once at the surface, the passenger domains of most autotransporters are folded into an elongated β-helix. In a cellular context, key molecules catalyze the assembly of the autotransporter β-barrel domain. However, how the passenger domain folds into its functional form is poorly u ...[more]

PMID: 29643377

Similar Datasets

Project description:BackgroundAutotransporters are attractive cell surface display vehicles as they lack complex adaptor proteins necessary for protein export. Recent reports have suggested that the native effector domain (? domain) and translocation domain (? domain) interact with each other to drive translocation of the effector domain to the outer membrane. In this report we compared the expression, surface localisation and folding of TEM-1 ?-lactamase (Bla) and maltose binding protein (MalE or MBP) fused to either full length Shigella flexneri IcsA (IcsA) autotransporter or to the ? domain alone (IcsA?) to determine the contribution of the native IcsA ? domain in presenting the fusion proteins on the surface of E. coli K-12 UT5600 (?ompT).ResultsExpression of IcsA-Bla was greater than IcsA?-Bla. High levels of IcsA-MalE were detected but IcsA?-MalE was not expressed. All fusion proteins other than IcsA?-MalE were localised to the outer membrane and were detected on the surface of UT5600 via immunofluorescence microscopy. All bacteria expressing IcsA-MalE were labelled with both ?-IcsA and ?-MBP. UT5600 expressing IcsA?-MalE was not labelled with ?-MBP. A third of UT5600 expressing IcsA-Bla were detectable with ?-Bla but only 5% of UT5600 (IcsA?-Bla) were labelled with ?-Bla. The correct folding of the Bla moiety when fused to IcsA and IcsA? was also retained as UT5600 expressing either fusion protein exhibited a decreased zone of inhibition in the presence of ampicillin. UT5600 expressing IcsA-Bla was more resistant compared to UT5600 expressing IcsA?-Bla.ConclusionsThe export mechanism of autotransporters is not well understood but accumulating evidence suggest a critical role for the native effector or ? domain in facilitating its own export via interactions with the translocation or ? domain. This is the first report directly comparing expression of heterologous proteins fused to the full length IcsA autotransporter and fusion to the ? domain alone. Protein expression and surface presentation of the fusion proteins were dramatically improved when fused to IcsA rather than IcsA?. Future studies involved in designing autotransporters as cell surface display vehicles would benefit from including the native ? domain. This work also provides further evidence for a key interaction between the autotransporter ? and ? domains.

Dataset Information

Molecular basis for the folding of ?-helical autotransporter passenger domains.

Publications

Molecular basis for the folding of β-helical autotransporter passenger domains.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets