| Abstract: | A procedure has been developed for the analysis of tyrosine O-sulfate in proteins. Samples are subjected to base hydrolysis with Ba(OH)2, neutralized with sulfuric acid, and the majority of other amino acids removed by chromatography on Dowex AG 50 X 8. The average recovery of tyrosine O-sulfate from these procedures was 43%. Tyrosine O-sulfate was identified by reverse-phase HPLC as the phenylthiocarbamyl derivative following precolumn derivatization with phenyl isothiocyanate. The method has been applied to bovine fibrinogen giving a tyrosine O-sulfate content ranging from 0.59 to 1.23 mol/mol. These procedures were also shown to be suitable for the analysis of the incorporation of [35S]sulfate into tyrosine O-sulfate residues in proteins by intact cells. |
