Ligand induced stabilization of the melting temperature of the HSV-1 single-strand DNA binding protein using the thermal shift assay
| Autor: | Kanchi Ravi Rupesh, Aaron Smith, Paul E. Boehmer |
|---|---|
| Rok vydání: | 2014 |
| Předmět: |
chemistry.chemical_classification
ICP8 Thermal shift assay Protein Stability Oligonucleotide viruses Biophysics Herpesvirus 1 Human Cell Biology biochemical phenomena metabolism and nutrition Ligands Ligand (biochemistry) Biochemistry Article DNA-Binding Proteins chemistry.chemical_compound chemistry Transition Temperature Nucleotide Molecular Biology DNA Binding Agent DNA Single-strand DNA-binding protein |
| Zdroj: | Biochemical and Biophysical Research Communications. 454:604-608 |
| ISSN: | 0006-291X |
| DOI: | 10.1016/j.bbrc.2014.10.145 |
| Popis: | We have adapted the thermal shift assay to measure the ligand binding properties of the herpes simplex virus-1 single-strand DNA binding protein, ICP8. By measuring SYPRO Orange fluorescence in microtiter plates using a fluorescence-enabled thermal cycler, we have quantified the effects of oligonucleotide ligands on the melting temperature of ICP8. We found that single-stranded oligomers raise the melting temperature of ICP8 in a length- and concentration-dependent manner, ranging from 1 °C for (dT)5 to a maximum of 9 °C with oligomers ≥10 nucleotides, with an apparent Kd of |
| Databáze: | OpenAIRE |
| Externí odkaz: |
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