High-throughput characterization of intrinsic disorder in proteins from the Protein Structure Initiative
| Autor: | Derrick E. Johnson, Vladimir N. Uversky, Tanguy Le Gall, Megan D. Sickmeier, Marc S. Cortese, Jingwei Meng, A. Keith Dunker, Bin Xue, Christopher J. Oldfield |
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| Rok vydání: | 2012 |
| Předmět: |
Models
Molecular Circular dichroism Knowledge Bases Proteolysis Crystal structure Biology Crystallography X-Ray Intrinsically disordered proteins Cleavage (embryo) Protein Structure Secondary Article chemistry.chemical_compound Structural Biology medicine Animals Humans Trypsin Databases Protein medicine.diagnostic_test Circular Dichroism Computational Biology Proteins Reference Standards Protein Structure Tertiary Biochemistry Myoglobin chemistry Structural Homology Protein Calibration Electrophoresis Polyacrylamide Gel Crystallization Protein Structure Initiative medicine.drug |
| Zdroj: | Journal of Structural Biology. 180:201-215 |
| ISSN: | 1047-8477 |
| DOI: | 10.1016/j.jsb.2012.05.013 |
| Popis: | The identification of intrinsically disordered proteins (IDPs) among the targets that fail to form satisfactory crystal structures in the Protein Structure Initiative represents a key to reducing the costs and time for determining three-dimensional structures of proteins. To help in this endeavor, several Protein Structure Initiative Centers were asked to send samples of both crystallizable proteins and proteins that failed to crystallize. The abundance of intrinsic disorder in these proteins was evaluated via computational analysis using predictors of natural disordered regions (PONDR®) and the potential cleavage sites and corresponding fragments were determined. Then, the target proteins were analyzed for intrinsic disorder by their resistance to limited proteolysis. The rates of tryptic digestion of sample target proteins were compared to those of lysozyme/myoglobin, apomyoglobin, and α-casein as standards of ordered, partially disordered and completely disordered proteins, respectively. At the next stage, the protein samples were subjected to both far-UV and near-UV circular dichroism (CD) analysis. For most of the samples, a good agreement between CD data, predictions of disorder and the rates of limited tryptic digestion was established. Further experimentation is being performed on a smaller subset of these samples in order to obtain more detailed information on the ordered/disordered nature of the proteins. |
| Databáze: | OpenAIRE |
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