Production of Ca-dependent high-affinity lectins with defined specificity by mutagenesis of PA-IIL lectin
| Autoři | |
|---|---|
| Rok publikování | 2011 |
| Druh | Konferenční abstrakty |
| Fakulta / Pracoviště MU | |
| Citace | |
| Popis | The bacterium Pseudomonas aeruginosa is a human opportunistic pathogen that can infect almost every human tissue in conditions of a lowered immunity barrier. The lectin PA IIL plays an important role in the bacteria’s virulence. It displays unusually high affinity for L fucose in the micromolar range. [1] This characteristic is correlated with the remarkable presence of two calcium ions in the binding site of the protein. In vitro mutagenesis was performed focusing on three single point mutants of PA-IIL in positions 22-23-24. The mutated amino acids belong to the “specificity-binding loop”. The structure of all mutants was determined by X-ray crystallography and their thermodynamic characterization was performed using isothermal titration calorimetry. The mutants were designed also by in silico methods, and the resulting structures were used for docking experiments. The in vitro mutagenesis in combination with computational methods allowed the key importance of amino acid 22 for the specificity of the lectin to be identified. |
| Související projekty: |