willis at gandalf.psf.sickkids.on.ca
Wed Apr 3 09:12:43 EST 1996
You don't tell us a lot about the protein or it's degradation products.
If they're small enough, gel filtration may be useful in separating the
full sized protein from the fragments but you will require a resin which
has a low molecular weight window of resolution.
It may be possible that there are differences in the interaction of the
proteolytic products with ion exchange resins (ie some of the charged
residues may have been clipped off)
Again, depending upon how much is lost in degradation, if your protein
has an affinity for some substrate, it may be that only the full sized
protein will bind to its ligand. This will allow you to use affinity
chromatography to isolate the full protein.
There may be differences in the solubility of the fragments in high salt
(ie ammonium sulfate) and you may be able to perform a salting out
experiment. If they are all equally soluble at the same levels of salt,
it may be possible to use this solution on a hydrophobic column and see
if there are differences in behaviour on this resin.
If there are differences in pI, it may be possible to perform
Good luck and let us know how it goes.
Randall C Willis, Aliquotes Press
"ALIQUOTES: A Journal of Molecular and Biochemical Humour"
58 Balfour Ave.
Toronto, ON M4C 1T6 CANADA
rogerb at microsoft.com willis at gandalf.psf.sickkids.on.ca
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