Directed Evolution and Engineering of Gallium-Binding Phage Clones-A Preliminary Study
| Autor: | Franziska L. Lederer, Robert Braun, Christina Zeitler, Sabine Matys, Katrin Pollmann, Nora Schönberger |
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| Rok vydání: | 2019 |
| Předmět: |
Biomedical Engineering
chemistry.chemical_element Bioengineering Peptide peptide structure 02 engineering and technology lcsh:Technology Biochemistry Article metal–peptide interaction Biomaterials 03 medical and health sciences Protein structure Gallium Site-directed mutagenesis Peptide sequence cysteine 030304 developmental biology chemistry.chemical_classification gallium 0303 health sciences lcsh:T Chemistry 021001 nanoscience & nanotechnology Directed evolution Combinatorial chemistry Amino acid phage surface display Molecular Medicine site-directed mutagenesis 0210 nano-technology Biotechnology Cysteine |
| Zdroj: | Biomimetics Biomimetics, Vol 4, Iss 2, p 35 (2019) Biomimetics 4(2019), 35 Volume 4 Issue 2 |
| ISSN: | 2313-7673 |
| Popis: | The phage surface display technology is a useful tool to screen and to extend the spectrum of metal-binding protein structures provided by nature. The directed evolution approach allows identifying specific peptide ligands for metals that are less abundant in the biosphere. Such peptides are attractive molecules in resource technology. For example, gallium-binding peptides could be applied to recover gallium from low concentrated industrial wastewater. In this study, we investigated the affinity and selectivity of five bacteriophage clones displaying different gallium-binding peptides towards gallium and arsenic in independent biosorption experiments. The displayed peptides were highly selective towards Ga3+ whereby long linear peptides showed a lower affinity and specificity than those with a more rigid structure. Cysteine scanning was performed to determine the relationship between secondary peptide structure and gallium sorption. By site-directed mutagenesis, the amino acids of a preselected peptide sequence are systematically replaced by cysteines. The resulting disulphide bridge considerably reduces the flexibility of linear peptides. Subsequent biosorption experiments carried out with the mutants obtained from cysteine scanning demonstrated, depending on the position of the cysteines in the peptide, either a considerable increase in the affinity of gallium compared to arsenic or an increase in the affinity for arsenic compared to gallium. This study shows the impressive effect on peptide&ndash target interaction based on peptide structure and amino acid position and composition via the newly established systematic cysteine scanning approach. |
| Databáze: | OpenAIRE |
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