I am,presently,working on ways to show the location of the sulfated
tyrosine in some mucin.It would be nice to narrow down to one peptide,which
has sulfated tyrosine.And,I
like amino acid analysis.The analysis is easy enough,since I am using the
AccQtag method
which is done at pHs where the sulfotyrosine is stable.I should be ok to do
HPLC separations at moderately acidic or neutral pH.The biggest impediment
has been in the
hydrolysis part,which is base hydrolysis.And,I think that I am getting
there.
As for proof,I do not know if sulfatases is sufficient.I,usually,treat the
resulting
sulfotyrosine with acid,for a very short time(6N HCl for 5min at 150).And,I
see only free
tyrosine.Sulfotyrosine elutes about 4 min earlier than tyrosine in the
AccQtag system.
Patrik ?nnerfjord <patrik.onnerfjord@medkem.lu.se>@aecom.yu.edu> on
05/31/2000 10:28:26 AM
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Subject: tyrosine sulfation of proteins?
Hi there,
I am working with MALDI-MS analysis of extracellular proteins which
in many cases are sulfated. Does anyone have experience in studying this?
Should one use sulfatases for the desulfation to prove that they are
sulfated or what is the general approach?
Thanks for all input!
/Patrik
Ph. D. Patrik ÷nnerfjord
Lund University, Dept. Cell and Molecular Biology
Section for Connective Tissue Biology
P.O. Box 94, 221 00 Lund
Fax: +46-46-211 3417 Phone: +46-46-222 3129
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