klenchin at REMOVE_TO_REPLY.facstaff.wisc.edu
Tue Jan 30 20:11:01 EST 2001
In article <3A768700.BE5D1F43 at slac.stanford.edu>, Mark Bowen
<mbo at slac.stanford.edu> wrote:
:Does anyone have experience with in vitro palimoylation? I've tried the
:reaction by adding palmitoyl CoA in DMSO to protein (25 kDa with 4 free
:cys) in the presence of 1% octylglucoside. This doesn't seems to be
:working based on SDS-PAGE.
Just out of curiosity - SNAP-25?
In any case, if this reaction proceeds with any reasonable rate
spontaneously, I am surprised, I did not know it. I was always
under impression that you need to have some source of palmitoyl
transferase activity. This certainly is the case for other lipid
modifications like myristoylation and geranyl geranylation.
If you express your protein as a fusion with something, reisolation
of it after in vitro palmitoylation in the presence of crude
extract should be relatively straightforward thing to do.
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