I am interested in phosphorylating my protein on specific sites. Are there any online tools to select a kinase that will specifically phosphorylate the sites of interest on my protein? What other resources are there available to determine the ideal kinases for my experiment?
You may try to use the online resource http://www.phosphosite.org which is a quite comprehensive tool. In case you are more interested in direct experimental data you may also want to have a look at the KinaseFinder service offered by ProQinase (http://www.proqinase.com/content/view/90).
Daniel
Motifscan is a free option where if u give your protein sequence, it gives you possible kinase substrate motif consensus.
http://scansite.mit.edu/motifscan_seq.phtml
Looking into the phosphosite based MS databases could also give insights into the context / stimuli/tissue where u naturally achieve these phosphorylations. This can also hint toward physiologically relevant kinases (for eg. Phosphorylation enriched in response to stress, in mitotic cells etc.). Prediction data together with this could help in making right conclusions
be carefull many kinases share in vitro the same consensus phosphorylation sequence
...AGC kinase are typical exemple...a RxxS motif is sufficient
an exception to this rule: lats-mob kinases!
You are approaching your question from the wrong direction. Try to establish the conditions under which your protein is phosphorylated and then pare down the possible protein kinases and test them in vitro (and with inhibitors, if available, in vivo). While there is overlapping specificity between a number of kinases, you really need to recapitulate what is going on in cells. Since you know the sequence around your "sites of interest", you could check out the NetworKIN resource to find likely kinases. However, our logic (and biology) is fallible so don't wed yourself to a specific kinase until you've established orthogonal data validating it. Good luck!
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