Laboratory of Toxicology, Faculty of Pharmacy, Osaka Ohtani University, 3-11-1 Nishikiori-kita, Tondabayashi, Osaka 584-8540, Japan.
Faculty of Pharmacy, Kindai University, 3-4-1 Kowakae, Higashi-osaka, Osaka 577-8502, Japan.
J Proteome Res. 2020 Aug 7;19(8):3033-3043. doi: 10.1021/acs.jproteome.0c00079. Epub 2020 May 29.
Prior investigations by our research group focused on the method development for the simultaneous analysis of sulfated and phosphorylated glycans. Herein, the developed method was applied to analyze minor acidic -glycans including sulfated and phosphorylated -glycans in human serum. First, 2-aminobenzoic acid-labeled minor acidic -glycans were enriched from the serum using a serotonin-immobilized column and were then separated into groups using hydrophilic interaction liquid chromatography, and analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Phosphorylated hybrid-type and sulfated bi-antennary -glycans were detected in the serum. In addition, we observed that multiple types of glucuronidated -glycans were present. These results indicate that the developed method is applicable to the analysis of glucuronidated as well as sulfated and phosphorylated -glycans. It was also applied to the sera obtained from 17 healthy subjects and 15 pancreatic cancer patients, and the profiles of sulfated, phosphorylated, and glucuronidated -glycans were compared. The expressed amount of glucuronidated -glycans was significantly decreased in some pancreatic cancer patients. Numerous examples of the -glycan analysis in human serum were reported, but phosphorylated and glucuronidated glycans were not investigated. The methods described herein allow the analysis of minor acidic glycans that are typically difficult to detect.
本研究组之前的研究重点是开发用于同时分析硫酸化和磷酸化聚糖的方法。在此，所开发的方法应用于分析包括人血清中的硫酸化和磷酸化在内的酸性聚糖。首先，使用 5-羟色胺固定化柱从血清中富集 2-氨基苯甲酸标记的酸性聚糖，然后使用亲水相互作用液相色谱将其分离成组，并通过基质辅助激光解吸/电离飞行时间质谱进行分析。在血清中检测到磷酸化杂合型和硫酸化双触角型聚糖。此外，我们观察到存在多种类型的硫酸化葡萄糖醛酸聚糖。这些结果表明，所开发的方法适用于分析硫酸化、磷酸化和葡萄糖醛酸化的聚糖。该方法还应用于 17 名健康受试者和 15 名胰腺癌患者的血清中，并对硫酸化、磷酸化和葡萄糖醛酸化聚糖的图谱进行了比较。在一些胰腺癌患者中，葡萄糖醛酸化聚糖的表达量显著降低。已报道了大量人血清中聚糖分析的实例，但未研究磷酸化和葡萄糖醛酸化聚糖。本文所述的方法允许分析通常难以检测的酸性聚糖。
