Conradi, Jens; Tegtmeyer, Nicole; Wozna, Marta; Wißbrock, Marco; Michalek, Carmela; Gagell, Corinna; Cover, Timothy L.; Frank, Ronald; Sewald, Norbert ; Backert, Steffen: An RGD helper sequence in CagL of Helicobacter pylori assists in interactions with integrins and injection of CagA. In: Frontiers in Cellular and Infection Microbiology. Jg.2. 2012, S. 70 ff.
Inhalt
- Human cell culture and H. pylori strains
- CagL protein expression and purification
- Site-directed mutagenesis
- Cell adhesion assays with WM-115 cells
- Helicobacter pylori binding to AGS cells
- CagL peptide synthesis
- CagL peptide SPOT arrays
- Circular dichroism spectroscopy
- Antibodies and Western Blotting
- Statistical analysis
- Results
- Synthetic RHS-cyclopeptides influence WM-115 cell adhesion to immobilized CagL mutants
- A linear RHS 15-mer peptide interferes with WM-115 adhesion to the CagLRAD and CagLRGA mutants
- The recombinant CagL mutants are correctly folded
- WM-115 cell adhesion studies with CagL RHS mutants
- Peptide array mapping of the CagL RHS motif for integrin binding
- Generation of RHS point mutations of CagL in H. pylori
- Mutation of the RHS motif inhibits CagA phosphorylation at early times of infection
- Mutation of the RHS motif exhibits significant signaling defects in H. pylori-induced ERK1/2 activation
- Mutation of the RHS motif results in reduced cortactin serine phosphorylation and AGS cell elongation
- Discussion
- Acknowledgments
- References