Differential Phosphorylation in response to a wild-type and mutant protein kinase

EK someone at microsoft.net
Fri Jan 9 12:13:52 EST 2004


> I think he means that he wants to show that the pattern of phosphorylation
> on the subtrate is different, ie that the mutant kinase has altered
> specificity. The general anti-pY/S/T antibodies won't reveal that; they'll
> only show whether the protein is phosphorylated at all, anywhere. So, if
> the WT kinase phosphprylates at S100 and S200, and the mutant only at
> S100, the general antibodies won't be a great deal of use (you ought to
> see a difference in the intensity of the band, but i'd be uneasy about
> trusting that, and it wouldn't give you positional information).
For some reason I was not aware that kinase could phosphorylate more that
one residue in a single protein/subunit. Still, if specificity of kinase
changes with mutation so that say S150 is not longer phosphorylated, and
Y250 is, then you should be able to see that with anti-pY/S.





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