Effects of Clostridium difficile Toxin A on the proteome of colonocytes studied by differential 2D electrophoresis

verfasst von

Johannes J. Zeiser, Jennifer Klodmann, Hans Peter Braun, Ralf Gerhard, Ingo Just, Andreas Pich

Abstract

Clostridium difficile is a spore-forming anaerobic pathogen, commonly associated with severe diarrhea or life-threatening pseudomembraneous colitis. Its main virulence factors are the single-chain, multi-domain toxin A (TcdA) and B (TcdB). Their glucosyltransferase domain selectively inactivates Rho proteins leading to a reorganization of the cytoskeleton. To study exclusively glucosyltransferase-dependent molecular effects of TcdA, human colonic cells (Caco-2) were treated with recombinant wild type TcdA and the glucosyltransferase deficient variant of the toxin, TcdA _gd for 24h. Changes in the protein pattern of the colonic cells were investigated by 2-D DIGE and LCMS/MS methodology combined with detailed proteome mapping. gdTcdA did not induce any detectable significant changes in the protein pattern. Comparing TcdA-treated cells with a control group revealed seven spots of higher and two of lower intensity (p<0.05). Three proteins are involved in the assembly of the cytoskeleton (β-actin, ezrin, and DPYL2) and four are involved in metabolism and/or oxidative stress response (ubiquitin, DHE3, MCCB, FABPL) and two in regulatory processes (FUBP1, AL1A1). These findings correlate well to known effects of TcdA like the reorganization of the cytoskeleton and stress the importance of Rho protein glucosylation for the pathogenic effects of TcdA.

Organisationseinheit(en)

Abteilung Pflanzenmolekularbiologie und Pflanzenproteomik
Institut für Pflanzengenetik

Externe Organisation(en)

Medizinische Hochschule Hannover (MHH)

Typ

Artikel

Journal

Journal of proteomics

Band

75

Seiten

469-479

Anzahl der Seiten

11

ISSN

1874-3919

Publikationsdatum

26.08.2011

Publikationsstatus

Veröffentlicht

Peer-reviewed

Ja

ASJC Scopus Sachgebiete

Biophysik, Biochemie

Ziele für nachhaltige Entwicklung

SDG 3 – Gute Gesundheit und Wohlergehen

Elektronische Version(en)

https://doi.org/10.1016/j.jprot.2011.08.012 (Zugang: Unbekannt)