Pore-forming Activity of the Escherichia coli Type III Secretion System Protein EspD.

The Journal of Biological Chemistry
Abhishek ChatterjeeArmando Jardim

Abstract

Enterohemorrhagic Escherichia coli is a causative agent of gastrointestinal and diarrheal diseases. Pathogenesis associated with enterohemorrhagic E. coli involves direct delivery of virulence factors from the bacteria into epithelial cell cytosol via a syringe-like organelle known as the type III secretion system. The type III secretion system protein EspD is a critical factor required for formation of a translocation pore on the host cell membrane. Here, we show that recombinant EspD spontaneously integrates into large unilamellar vesicle (LUV) lipid bilayers; however, pore formation required incorporation of anionic phospholipids such as phosphatidylserine and an acidic pH. Leakage assays performed with fluorescent dextrans confirmed that EspD formed a structure with an inner diameter of ∼2.5 nm. Protease mapping indicated that the two transmembrane helical hairpin of EspD penetrated the lipid layer positioning the N- and C-terminal domains on the extralumenal surface of LUVs. Finally, a combination of glutaraldehyde cross-linking and rate zonal centrifugation suggested that EspD in LUV membranes forms an ∼280-320-kDa oligomeric structure consisting of ∼6-7 subunits.

References

May 24, 1991·Science·A LupasJ Stock
Aug 15, 1995·Proceedings of the National Academy of Sciences of the United States of America·K G JarvisJ B Kaper
Feb 28, 1995·Proceedings of the National Academy of Sciences of the United States of America·T K McDanielJ B Kaper
Nov 1, 1995·Protein Science : a Publication of the Protein Society·C N PaceT Gray
Apr 1, 1996·Protein Science : a Publication of the Protein Society·P KaulR J Collier
Feb 27, 1999·Current Opinion in Microbiology·R DeVinneyB B Finlay
Dec 5, 2000·Current Biology : CB·C M RosenbergerB B Finlay
Sep 26, 2001·Applied and Environmental Microbiology·P MarteauG Corthier
Feb 28, 2002·Journal of Molecular Biology·Jessica P DawsonDonald M Engelman
Jun 19, 2002·Molecular Microbiology·Emma J McGhieVassilis Koronakis
Feb 14, 2003·Lancet·Francisco Guarner, Juan-R Malagelada
Jun 5, 2003·Cellular Microbiology·Jean-Philippe NougayrèdeMichael S Donnenberg
Jun 28, 2003·Molecular Microbiology·Sarah J DaniellGad Frankel
Mar 26, 2004·Nature Reviews. Microbiology·James B KaperHarry L Mobley
Nov 25, 2004·Progress in Biophysics and Molecular Biology·Michael W Parker, Susanne C Feil
Dec 4, 2004·Cell Death and Differentiation·J K Crane, C M Vezina

❮ Previous
Next ❯

Citations

Mar 26, 2016·Frontiers in Immunology·Julie Guignot, Guy Tran Van Nhieu
Nov 8, 2016·Frontiers in Cellular and Infection Microbiology·Meztlli O GaytánBertha González-Pedrajo
May 26, 2018·FEMS Microbiology Letters·Biao YuanSpyridoula Karamanou
Jun 3, 2021·Microorganisms·Miguel Díaz-GuerreroBertha González-Pedrajo

❮ Previous
Next ❯

Related Concepts

Related Feeds

ASBMB Publications

The American Society for Biochemistry and Molecular Biology (ASBMB) includes the Journal of Biological Chemistry, Molecular & Cellular Proteomics, and the Journal of Lipid Research. Discover the latest research from ASBMB here.