Purification of metal-dependent lysine deacetylases with consistently high activity
| Autor: | Chanel D. Perry, Richard G. Painter, Ashley N. Matthew, Kyara A. Nichols, Asia N. Matthew-Onabanjo, Rashad A. Haynes, Terry J. Watt, Tasha B. Toro, Elena Y. Glotser, Derek R. Bratcher, Melyssa R. Bratton, Jenae R. Bryant |
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| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Lysine Biology Histone Deacetylases Article Metal 03 medical and health sciences Affinity chromatography Escherichia coli Sf9 Cells Animals Humans High activity chemistry.chemical_classification 030102 biochemistry & molecular biology Circular Dichroism Substrate (chemistry) Cobalt Recombinant Proteins Repressor Proteins 030104 developmental biology Enzyme chemistry Biochemistry Acetylation visual_art visual_art.visual_art_medium Electrophoresis Polyacrylamide Gel Histone deacetylase Biotechnology |
| Zdroj: | Protein Expression and Purification. 141:1-6 |
| ISSN: | 1046-5928 |
| Popis: | Metal-dependent lysine deacetylases (KDACs) are involved in regulation of numerous biological and disease processes through control of post-translational acetylation. Characterization of KDAC activity and substrate identification is complicated by inconsistent activity of prepared enzyme and a range of multi-step purifications. We describe a simplified protocol based on two-step affinity chromatography. The purification method is appropriate for use regardless of expression host, and we demonstrate purification of several representative members of the KDAC family as well as a selection of mutated variants. The purified proteins are highly active and consistent across preparations. |
| Databáze: | OpenAIRE |
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