通过反相高效液相色谱法鉴定蛋白质中的酪氨酸O-硫酸盐：碱水解结合异硫氰酸苯酯柱前衍生化的应用。
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.
作者信息
Christie D L, Hill R M, Isakow K, Barling P M
出版信息
Anal Biochem. 1986 Apr;154(1):92-9. doi: 10.1016/0003-2697(86)90500-2.
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.
已开发出一种分析蛋白质中酪氨酸 O-硫酸盐的方法。样品用 Ba(OH)₂ 进行碱水解，用硫酸中和，然后通过 Dowex AG 50 X 8 柱色谱去除大多数其他氨基酸。这些方法中酪氨酸 O-硫酸盐的平均回收率为 43%。用异硫氰酸苯酯进行柱前衍生后，通过反相高效液相色谱将酪氨酸 O-硫酸盐鉴定为苯硫基甲酰胺衍生物。该方法已应用于牛纤维蛋白原，其酪氨酸 O-硫酸盐含量范围为 0.59 至 1.23 mol/mol。这些方法还被证明适用于分析完整细胞将 [³⁵S]硫酸盐掺入蛋白质中酪氨酸 O-硫酸盐残基的情况。
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