Mutagenising phosphorylation sites
drm21 at mole.bio.cam.ac.uk
Tue Aug 1 09:15:14 EST 1995
I wonder if someone out there could give me some advice about mutagenising
Serine residues suspected of being phosphorylation sites.
As far as I can gather, it is common to change the Serine to an Alanine to
mimic the always-unphosphorylated state, and to an Aspartate to mimic
Is this correct? Does anyone know why this is done rather than Serine->
Cysteine, which should be both unphosphorylatable and (presumably - I'm
not a great chemist) more chemically similar to Serine? (This is an
intracellular protein, so formation of disulfide bonds shouldn't be a
Similarly, a naive look at the structures of phosphoserine, aspartate and
glutamate suggest to me that glutamate is more like phosphoserine than
aspartate is. Am I mistaken?
Any guidance on why Ala and Asp should be used instead of Cys and Glu (or vice
versa) would be much appreciated.
Thanks for any help!
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