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How did I think when I prepare FLIM PIP3 PSD95?

At first, I reffered the paper

At first, N-terminus is very important for target PSD-95 to the membrane.


[[PSD-95-01>http://www.jbc.org/content/273/41/26269.full ]]

[[PSD-95-02>http://www.nature.com/nrn/journal/v5/n10/pdf/nrn1517.pdf]]

[[PSD-95-03>http://www.jbc.org/content/277/5/3640.full]]

How did I think when I prepare FLIM PIP3 PSD95?

At first, I refered to following paper.
[[PSD-95>http://www.jbc.org/content/274/1/532.short]]

I wanted a targeting domain as less as possible.
N-segment-PDZ1-PDZ2 protein is enough for this protein targeting to membrane, multimer, Kv1.4 channel binding and clustering, indicating that PDZ3 is not important for membrane targeting.

the fusion protein in which GFP is attached to PDZ1-PDZ2 domain from C-terminus seems to give this protein tergeting to the membrane.
But I don't want to do my probe to multimer.
And It takes much of time to use C-terminus version of my probe.
And I thought that only N-segment is not enough to terget to the spines.

So I fused my probe to the left side of N-segment-PDZ1-PDZ2.

 
N-terminus is very important for target PSD-95 to the membrane.
1. terget domain

[[PSD-95-01>http://www.jbc.org/content/273/41/26269.full ]]

[[PSD-95-02>http://www.nature.com/nrn/journal/v5/n10/pdf/nrn1517.pdf]]

[[PSD-95-03>http://www.jbc.org/content/277/5/3640.full]]