Due to the knowledge that one protein kinase have many Intracellular target proteins, for making the experiment, are there any methods that make protein kinase enzyme phosphorylates only one desired target protein? Or there are any strategies to make sure that protein kinase phosphorylates only desired target in the experiment to ensure the reliability of results from phosphorylation study?
Thank you in advance for your answer.
@Adam B Shapiro Thank you very much for the answer. Please let me ask you some another question. Is this technique can used for the experiment on cell culture? For the previously question, I had a plan in study the effect of phosphorylation on some cell activities. And when I tell my seniors, they asked me that how to make sure that the results is from the phosphorylation of targeted protein only, not from other proteins which is a substrate for the kinase use in the study (I plan to choose PKA for the phosphorylation) And I really has no idea about this question. Thank you in advance for your help!
One uses purified proteins to study the specific interaction between them. This method is not relevant to cells.
In cells, you could study the effect on one specific protein by site-directed mutagenesis of the gene for that protein. Individual sites of phosphorylation are mutated to acidic residues to mimic permanent phosphorylation, or to alanine to prevent phosphorylation.
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