Hello. I want to crystallize kinase domain of a protein in active form. The active form of the domain has specifically phosphorylated tyrosine residues. However, I am not sure if I can do so in a bacterial expression system. Apart from this is it possible to phosphorylate specific tyrosine residues (if so it would be kind enough if one can share any protocol/literature)?
U cannot get PTM in E. coli, try using different expression system like yeast or baculovirus.
Bacteria will not do it. You may use a recombinant enzyme to do it on a purified protein and separate components later.
Timir Tripathi Thank you, but I am still not sure if the PTM or in this case phosphorylation will be on specfic tyrosines only.
Wieslaw Swietnicki can you provide literature similar to the same.
Look up kinases and their specificities. Some are non-specific and other require a specific substrate sequence. Without knowing anything about your enzyme, there is no answer to your question.
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