Identification of tyrosine O-sulfate in proteins by reverse-phase high-performance liquid chromatography: use of base hydrolysis combined with precolumn derivatization using phenyl isothiocyanate.
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.
D L
DL
Christie
R M
RM
Hill
K
K
Isakow
P M
PM
Barling
D000818:Animals; D001464:Barium; D017609:Barium Compounds; D002417:Cattle; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005340:Fibrinogen / Q000032:analysis; D006868:Hydrolysis; D017879:Isothiocyanates; D011506:Proteins / Q000032:analysis*; D013861:Thiocyanates; D014443:Tyrosine / Q000031:analogs & derivatives* / Q000032:analysis
D017609:Barium Compounds; D017879:Isothiocyanates; D011506:Proteins; D013861:Thiocyanates; C005441:phenylisothiocyanate; C012766:barium hydroxide; D001464:Barium; C005662:tyrosine O-sulfate; D014443:Tyrosine; D005340:Fibrinogen