Details
| Original language | English |
|---|---|
| Pages (from-to) | 469-479 |
| Number of pages | 11 |
| Journal | Journal of proteomics |
| Volume | 75 |
| Issue number | 2 |
| Publication status | Published - 26 Aug 2011 |
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.
Keywords
- 2D-DIGE, Caco-2, Clostridium difficile, Proteome map, Toxin A
ASJC Scopus subject areas
- Biochemistry, Genetics and Molecular Biology(all)
- Biophysics
- Biochemistry, Genetics and Molecular Biology(all)
- Biochemistry
Sustainable Development Goals
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In: Journal of proteomics, Vol. 75, No. 2, 26.08.2011, p. 469-479.
Research output: Contribution to journal › Article › Research › peer review
}
TY - JOUR
T1 - Effects of Clostridium difficile Toxin A on the proteome of colonocytes studied by differential 2D electrophoresis
AU - Zeiser, Johannes J.
AU - Klodmann, Jennifer
AU - Braun, Hans Peter
AU - Gerhard, Ralf
AU - Just, Ingo
AU - Pich, Andreas
N1 - Funding information: This work was supported by a grant to AP and IJ from the Deutsche Forschungsgemeinschaft ( Pi278/2-2 ) and to RG by the Deutsche Forschungsgemeinschaft ( SFB621 , project B5). The authors thank Helma Tatge, Angelika Neumann and Dagmar Lewejohann for excellent technical assistance and Christina Rode for help with Delta2D.
PY - 2011/8/26
Y1 - 2011/8/26
N2 - 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.
AB - 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.
KW - 2D-DIGE
KW - Caco-2
KW - Clostridium difficile
KW - Proteome map
KW - Toxin A
UR - http://www.scopus.com/inward/record.url?scp=82355173202&partnerID=8YFLogxK
U2 - 10.1016/j.jprot.2011.08.012
DO - 10.1016/j.jprot.2011.08.012
M3 - Article
C2 - 21890007
AN - SCOPUS:82355173202
VL - 75
SP - 469
EP - 479
JO - Journal of proteomics
JF - Journal of proteomics
SN - 1874-3919
IS - 2
ER -