Folding of a bacterial integral outer membrane protein is initiated in the periplasm

Nature Communications
Rakesh SikdarHarris D Bernstein

Abstract

The Bam complex promotes the insertion of β-barrel proteins into the bacterial outer membrane, but it is unclear whether it threads β-strands into the lipid bilayer in a stepwise fashion or catalyzes the insertion of pre-folded substrates. Here, to distinguish between these two possibilities, we analyze the biogenesis of UpaG, a trimeric autotransporter adhesin (TAA). TAAs consist of three identical subunits that together form a single β-barrel domain and an extracellular coiled-coil ("passenger") domain. Using site-specific photocrosslinking to obtain spatial and temporal insights into UpaG assembly, we show that UpaG β-barrel segments fold into a trimeric structure in the periplasm that persists until the termination of passenger-domain translocation. In addition to obtaining evidence that at least some β-barrel proteins begin to fold before they interact with the Bam complex, we identify several discrete steps in the assembly of a poorly characterized class of virulence factors.

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Citations

Jan 20, 2018·Science·Alexandra I C HöhrNils Wiedemann
Aug 15, 2018·Molecular Microbiology·Janine H PetersonHarris D Bernstein
Aug 9, 2018·Proceedings of the National Academy of Sciences of the United States of America·Karl LundquistJames C Gumbart
Jun 13, 2018·FEMS Microbiology Letters·Guillaume Mas, Sebastian Hiller
Dec 23, 2019·Medical Microbiology and Immunology·Andreas R KiesslingAdrian Goldman
May 5, 2018·FEMS Microbiology Letters·David RanavaRaffaele Ieva
Jul 28, 2019·Nature Communications·Matthew T Doyle, Harris D Bernstein
Aug 31, 2020·MicrobiologyOpen·Hawzeen S KhalilJack C Leo
Feb 10, 2021·Current Opinion in Microbiology·David Tomasek, Daniel Kahne
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Apr 27, 2021·Current Opinion in Structural Biology·Jérémy Guérin, Susan K Buchanan
Mar 7, 2019·EcoSal Plus·Harris D Bernstein
Jun 3, 2021·Toxins·Nathalie Dautin

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Methods Mentioned

BETA
immunoprecipitations
immunoprecipitation
irradiated
protein folding
PCR

Software Mentioned

MaxQuant
UCSF Chimera

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