)Hi all,
I tried to inhibit pSRC downstream signalling by using SRC kinase inhibitor PP2. In my experiments I could detect a reduction in protein levels downstream SRC, However, SRC phosphorylation levels were increasing upon inhibition.
Did anybody saw such effect when using PP2? or is there any explanation for this?
Thanks in advance.
Eddy Breemen
Maybe our functional assay, that measures kinase activity in an incubated lysate, could give additional insights. Indeed we also see that pressense of a kinase does not equal activity. You can learn more about us at www.pamgene.com, or send me an email at [email protected]. Looking forward!
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Becky Tu-Sekine
Hi Fatima,
- I have also observed this in a MEF cell line variant that has low basal Src activity. In my case I found a dose dependent increase in Y416 Src (0.5uM to 5 uM PP2 over weekend - approx 60 hours). A recent paper found that several non-specific Src family inhibitors (including PP2) seem to have this effect by causing a conformational change that leads to the association of Src with FAK. Once the inhibitor is displaced FAK can then phosphorylate Src (see PMID: 33761359, Higuchi et al 2021). In other words, Carol A Heckman may have hit the nail on the head! I'm not sure if this is what is happening in my cells or yours, but it gives a reasonable explanation. I am moving away from PP2 since it doesn't really target Src very well (it's listed as a Lck/Fyn inhibitor). Maybe try dasatinib or SU6656? Good luck!
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