Cloned (Comment) | Organism |
---|---|
p38alpha active mutants subcloned into vector pET-28a and expressed in Rosetta strain of Escherichia coli | Mus musculus |
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 | 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 | UniProt | Comment | Textmining |
---|---|---|---|
Mus musculus | - |
- |
- |
Purification (Comment) | Organism |
---|---|
p38alpha active mutants, by gel filtration, on Ni2+-column | Mus musculus |
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 | Comment | Organism |
---|---|---|
p38 mitogen-activated protein kinase | - |
Mus musculus |
p38a MAP kinase | - |
Mus musculus |
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 |