A ClpS-based N-terminal amino acid binding reagent with improved thermostability and selectivity
| Autor: | Jennifer Tullman, Zvi Kelman, John P. Marino, Makenzie Christensen |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
0106 biological sciences
chemistry.chemical_classification 0303 health sciences Environmental Engineering Chemistry Biomedical Engineering Bioengineering Peptide Context (language use) 01 natural sciences Combinatorial chemistry Amino acid 03 medical and health sciences Residue (chemistry) Protein sequencing 010608 biotechnology Reagent Amino acid binding 030304 developmental biology Biotechnology Thermostability |
| Zdroj: | Biochemical Engineering Journal. 154:107438 |
| ISSN: | 1369-703X |
| DOI: | 10.1016/j.bej.2019.107438 |
| Popis: | Realization of binding reagents that can perform high-fidelity, sequential recognition and detection of amino acids is important for the success of many proposed approaches for single-molecule protein sequencing. Towards this purpose, a variant of the Agrobacterium tumefaciens protein ClpS was previously engineered with improved binding affinity for phenylalanine at the N-terminus of a peptide. In this study, this variant was further engineered for attributes necessary for robust application as a biotechnology reagent such as increased thermostability and improved selectivity under different buffer conditions. The stabilized variant was further characterized in terms of selectivity for the N-terminal residue in the context of different peptides with varying residues at positions 2 (P2) and 3 (P3). The implication of the study for the use of ClpS based reagents in peptide sequencing technologies are described. |
| Databáze: | OpenAIRE |
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