Comparative Glycomic Analysis of Sialyl Linkage Isomers by Sialic Acid Linkage-Specific Alkylamidation in Combination with Stable Isotope Labeling of α2,3-Linked Sialic Acid Residues
Comparative Glycomic Analysis of Sialyl Linkage Isomers by Sialic Acid Linkage-Specific Alkylamidation in Combination with Stable Isotope Labeling of α2,3-Linked Sialic Acid Residues
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DOI：
10.1021/acs.analchem.9b03617
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发表时间：
2019-11-05
影响因子：
7.4
通讯作者：
Sakamoto, Naoya
中科院分区：
文献类型：
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作者：
Hanamatsu, Hisatoshi;Nishikaze, Takashi;Sakamoto, Naoya
唾液酸通过α 2，3、α 2，6或α 2，8键形成糖蛋白上聚糖链中的末端糖，并且唾液酸键的结构异构体在细胞识别和其他生理过程中发挥各种功能作用。我们最近开发了一种新的程序的基础上唾液酸键特异性烷基酰胺化通过内酯开环（氨解-SALSA）。在此，我们研究了使用胺盐酸盐的α 2，3-连接的唾液酸残基（iSALSA）的同位素标记。使用胺盐酸盐的SALSA的一个限制可以通过仅添加叔丁胺（t-BA）作为酸清除剂来解决，并且可以使用iSALSA进行比较和定量糖组学分析。我们还开发了定量糖组学分析，使用双同位素标记的聚糖，通过在还原和非还原端分别用氨氧基官能化的双烷基精氨酸甲酯（aoWR）和iSALSA进行衍生化。此外，我们证明，在肝纤维化过程中，血清中的α 2，3-连接唾液酸聚糖的量改变，使用基质辅助激光解吸电离飞行时间质谱（MALDI-TOF MS）和液相色谱MS（LC/MS）分析。我们发现A3（3，6，6）与A3 F（3，6，6）的比值随着肝纤维化的进展而沿着逐渐降低。因此，这些聚糖改变是非酒精性脂肪性肝炎（NASH）纤维化进展的潜在诊断标志物。
Sialic acids form the terminal sugars in glycan chains on glycoproteins via alpha 2,3, alpha 2,6, or alpha 2,8 linkages, and structural isomers of sialyl linkages play various functional roles in cell recognition and other physiological processes. We recently developed a novel procedure based on sialic acid linkage-specific alkylamidation via lactone ring opening (aminolysis-SALSA). Herein, we have investigated an isotope labeling of alpha 2,3-linked sialic acid residues (iSALSA) using amine hydrochloride salts. One limitation of SALSA using amine hydrochloride salts may be solved by adding only tert-butylamine (t-BA) as an acid scavenger, and comparative and quantitative glycomic analyses can be performed using iSALSA. We also developed quantitative glycomic analysis using dual isotope-labeled glycans by derivatizing with aminooxy-functionalized tryptophanylarginine methyl ester (aoWR) and iSALSA at the reducing and nonreducing end, respectively. Furthermore, we demonstrate that the amount of alpha 2,3-linked sialoglycans in serum are altered during liver fibrosis using matrix-assisted laser desorption ionization-time-of-flight mass spectrometry (MALDI-TOF MS) and liquid chromatography MS (LC/MS) analyses. We revealed that the ratio of A3(3,6,6) to A3F(3,6,6) was gradually decreased along with liver fibrosis progression. Therefore, these glycan alterations are potential diagnostic markers of nonalcoholic steatohepatitis (NASH) fibrosis progression.