Influence of different N- and O-linked carbohydrates on the retention times of synthetic peptides in reversed-phase high-performance liquid chromatography.
Influence of different N- and O-linked carbohydrates on the retention times of synthetic peptides in reversed-phase high-performance liquid chromatography.
复制标题
不同N-和O-连接碳水化合物对合成肽在反相高效液相色谱中保留时间的影响。
DOI：
10.1016/0021-9673(92)85457-5
复制
发表时间：
1992
期刊：
影响因子：
--
通讯作者：
Thurin,J
中科院分区：
文献类型：
--
作者：
OtvosJr,L;Urge,L;Thurin,J
在固相上合成了由6-19个氨基酸残基和不同的单糖和二糖连接到单个天冬酰胺和丝氨酸残基组成的糖肽，并通过反相高效液相色谱和圆二色谱进行了表征。结果表明，由于糖基化导致的保留时间缩短可能与糖部分长度增加相关。相同序列的磷酸化比单糖的糖基化减少了1.6倍的保留时间。对N-糖肽合成的结构单元天冬酰胺衍生化和糖基化时，与柱的结合取决于二糖的结构。然而，这种洗脱时间的结构依赖性在最终糖肽中消失。虽然糖基化和磷酸化都导致肽骨架的二级结构改变，但保留时间似乎反映了亲水性的增加比键合相表面上诱导的构象取向更强烈。
Glycopeptides consisting of 6–19 amino acid residues and different mono- and disaccharides attached to single asparagine and serine residues were synthesized on solid-phase and were characterized by reversed-phase high-performance liquid chromatography and circular dichroism. It was shown that the decreased retention times due to glycosylation could be correlated with the increasing length of the sugar moiety. Phosphorylation of the same sequences reduced the retention times 1.6 times more than glycosylation with monosaccharides did. The binding to the column was dependent on the structure of the disaccharide when derivatized and glycosylated asparagine, the building block of N-glycopeptide syntheses was studied. However, this structural dependence of the elution times disappeared in the final glycopeptides. Although both glycosylation and phosphorylation resulted in altered secondary structure of the peptide backbone, it appears that the retention times reflect the increased hydrophilicity more strongly than induced conformational orientation on the surface of the bonded phase.