Hydroxylamine for cleaving proteins?
pd1 at mole.bio.cam.ac.uk
Thu Jan 18 15:38:42 EST 1996
Alan Kay wrote:
> marciam at unixg.ubc.ca (Marcia MacDonald) wrote:
> >I am hoping to use hydroxylamine to cleave a phosphorylated protein but
> >I have been unable to find any recent references on the procedure. Has
> >anyoiaelse used this method? If so, will it interfere with the
> >phosphorylated residues? Any information at all would be helpful.
> I have used hydroxylamine a lot to cleave off the fusion part of
> recombinant fusion proteins. The preferred cleavage site of
> hydroxylamine is asparagine-glycine. According to "Methods in
> Enzymology" (probably volume 47, quite old) it is thought that
> the side chain of the asparagine lies over the glycine and at
> basic pH there is cyclisation and cleavage by hydroxylamine.
> Whether phosphorylation of ser or thr in an asn-gly-ser or thr
> context will affect this, I do not know.You must stay under
> pH 10, because this rearranges the asparagine.
It is Vol 47, the ref is;
Bornstein and Balian (1977). Methods Enzymol. 47, p132-
Division of Virology
Department of Pathology
University of Cambridge
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