Comparative and optimized dabsyl-amino acid analysis of synthetic phosphopeptides and glycopeptides.
Comparative and optimized dabsyl-amino acid analysis of synthetic phosphopeptides and glycopeptides.
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合成磷酸肽和糖肽的比较和优化 dabsyl 氨基酸分析。
DOI：
10.1016/0021-9673(94)80459-1
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发表时间：
1994
期刊：
影响因子：
--
通讯作者：
OtvosJr,L
中科院分区：
文献类型：
--
作者：
Gorbics,L;Urge,L;OtvosJr,L
采用Dabsyl-Cl衍生化和反相高效液相色谱分离技术，研究了N-乙酰氨基葡萄糖、N-乙酰氨基葡萄糖和N-乙酰氨基半乳糖糖肽氨基酸分析的最佳条件。通过比较三种不同色谱柱上的二氨基磷酸和二氨基糖的层析行为，发现这两种修饰的dabsyl衍生物与柱结合的机理是不同的。研究了糖基和磷酸基团的酸敏性。我们发现，虽然磷酸肽分析的最佳水解条件与多肽序列有关，但通常存在一个适用的条件，以获得最高的糖肽回收率。大多数糖肽的气相水解时间以1h为宜，磷酸肽的气相水解时间以1.5h为宜。当N-乙酰氨基葡萄糖和N-乙酰半乳糖胺作为糖肽的双糖侧链的一部分被结合时，分析被扩展到成功地验证了这些糖的存在。
The optimal conditions for amino acid analysis of phosphopeptides and N-acetylglucosamine- and N-acetylgalactosamine-containing glycopeptides were investigated by dabsyl-Cl derivatization and reversed-phase high-performance liquid chromatographic separation. By comparing the chromatographic behaviour of the dabsylated phosphoamino acids and dabsylated aminosugars on three different columns, it appears that the mechanism of binding to the column is different for the two modified dabsyl derivatives. The acid sensitivities of sugar and phosphate groups were also investigated. We found that while the optimal hydrolysis conditions for phosphopeptide analysis are peptide sequence-dependent, there is generally an applicable condition for the highest recovery of glycopeptides. A 1-h gas-phase hydrolysis time seems to be appropriate for the majority of glycopeptides and 1.5 h is suitable for the majority of phosphopeptides. The analysis was extended to the successful verification of the presence of the N-acetylglucosamine and the N-acetylgalactosamine moieties when these sugars were incorporated as parts of a disaccharide side chain of glycopeptides.