Mammal hyaluronidase activity on chondroitin sulfate and dermatan sulfate: Mass spectrometry analysis of oligosaccharide products
Mammal hyaluronidase activity on chondroitin sulfate and dermatan sulfate: Mass spectrometry analysis of oligosaccharide products
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DOI：
10.1093/glycob/cwab004
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发表时间：
2021-02-17
期刊：
影响因子：
4.3
通讯作者：
Daniel, Regis
中科院分区：
文献类型：
--
作者：
Bilong, Melanie;Bayat, Parisa;Daniel, Regis
哺乳动物透明质酸酶是参与透明质酸（HA）催化的内切-N-乙酰基-D-氨基己糖苷酶，但它们在硫酸软骨素（CS）催化中的作用也被研究。HA和CS是参与多种生理和病理过程的糖胺聚糖，了解它们的代谢具有重要意义。以前已经报道了在透明质酸酶作用下CS降解的数据，但仍有必要对CS解聚产物进行详细的结构研究，以提高我们对透明质酸酶CS解聚活性的认识。为此目的，已经通过高分辨率Orbitrap质谱（MS）和极紫外（XUV）光解离串联MS对在透明质酸酶对各种CS亚型进行酶促解聚后形成的CS寡糖进行了精细结构表征。精确的质量测量显示在消化CS-A和CS-C后形成宽尺寸范围的均匀寡糖，在主要消化产物中，有少量的奇数寡糖形成，而在CS-B上未检测到透明质酸酶活性。此外，在CS-A和CS-C的消化混合物中的寡糖分布中观察到轻微差异，对于CS-C，较长寡糖的贡献显著更高。通过XUV光解离实验确定的CS寡糖产物的序列验证了哺乳动物透明质酸酶催化的选择性β（1 -> 4）糖苷键断裂。哺乳动物透明质酸酶产生六糖和更高寡糖的能力支持其在锚定至膜蛋白聚糖的CS的催化剂中和在细胞外基质中的作用。
Mammalian hyaluronidases are endo-N-acetyl-D-hexosaminidases involved in the catabolism of hyaluronic acid (HA) but their role in the catabolism of chondroitin sulfate (CS) is also examined. HA and CS are glycosaminoglycans implicated in several physiological and pathological processes, and understanding their metabolism is of significant importance. Data have been previously reported on the degradation of CS under the action of hyaluronidase, yet a detailed structural investigation of CS depolymerization products remains necessary to improve our knowledge of the CS depolymerizing activity of hyaluronidase. For that purpose, the fine structural characterization of CS oligosaccharides formed upon the enzymatic depolymerization of various CS subtypes by hyaluronidase has been carried out by high-resolution Orbitrap mass spectrometry (MS) and extreme UV (XUV) photodissociation tandem MS. The exact mass measurements show the formation of wide size range of even oligosaccharides upon digestion of CS-A and CS-C comprising hexa- and octa-saccharides among the main digestion products, as well as formation of small quantities of odd-numbered oligosaccharides, while no hyaluronidase activity was detected on CS-B. In addition, slight differences have been observed in the distribution of oligosaccharides in the digestion mixture of CS-A and CS-C, the contribution of longer oligosaccharides being significantly higher for CS-C. The sequence of CS oligosaccharide products determined by XUV photodissociation experiments verifies the selective beta(1 -> 4) glycosidic bond cleavage catalyzed by mammal hyaluronidase. The ability of the mammal hyaluronidase to produce hexa- and higher oligosaccharides supports its role in the catabolism of CS anchored to membrane proteoglycans and in extra-cellular matrix.