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Literature summary for 2.7.11.24 extracted from
- High-resolution diffracting crystals of intrinsically active p38alpha MAP kinase: a case study for low-throughput approaches (2007), Acta Crystallogr. Sect. D, 63, 260-265.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| p38alpha active mutants subcloned into vector pET-28a and expressed in Rosetta strain of Escherichia coli | Mus musculus |
Crystallization (Commentary)
| Crystallization (Comment) | Organism |
|---|---|
| p38alpha active mutants crystallized by sitting-drop vapour-diffusion method, mutant D176A/F327L to 1.45 A resolution, crystals of the three p38alpha mutants belong to the orthorhombic space group P212121, with one molecule in the asymmetric unit | Mus musculus |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| D176A | active p38alpha mutant, crystals do not appear spontaneously, cross-seeding approaches using crystals of mutant D176A+F327L as the source of microseeds results in crystals suitable for X-ray analysis | Mus musculus |
| D176A/F327L | most active p38alpha mutant, improvement of crystallization assays, obtained with abolishing phosphorylation completely by reducing both the temperature and duration of induction and by significantly shortening the N-terminal hexahistidine spacer, facilitating the growth of well diffracting crystals | Mus musculus |
| D176A/F327S | active p38alpha mutant, crystals do not appear spontaneously, cross-seeding approaches using crystals of mutant D176A+F327L as the source of microseeds results in crystals suitable for X-ray analysis | Mus musculus |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Mus musculus | - |
- |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| p38alpha active mutants, by gel filtration, on Ni2+-column | Mus musculus |
Storage Stability
| Storage Stability | Organism |
|---|---|
| -80°C, 100 mM NaCl, 50 mM Tris-HCl buffer, pH 7.4, 10 mM DTT, 10 mM MgCl2, 5% glycerol | Mus musculus |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| p38 mitogen-activated protein kinase | - |
Mus musculus |
| p38a MAP kinase | - |
Mus musculus |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| additional information | - |
using a relatively low induction temperature (21°C in comparison to 32°C) phosphorylation is almost completely prevented. Combining a short 5 h induction with a low expression temperature (21°C) results in highly homogeneous unphosphorylated protein | Mus musculus |
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Release 2023.1 (January 2023)
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