In this paper, when GFP and RFP or DsRED were expressed to the membrane using palmitoyl targeting domain, FLIM value becomes shorter, which means intermolecular FLIM occurs. This is the case in culture cells.
I have no idea that in spines this intermolecular FRET occurs. However a spine is smaller space than culture cells. So we can predict that such a FRET is gonna happen in spines.
preparation of point mutations of PH domain of PIP3FLIM.
My probe is attached at the membrane.
So possibly binding of PH domain to PIP3 is more than normal state.
So I make the binind ability of PH domain weak.
The point mutations I try to target are as follows.
GTG AAG ACC