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从牛角膜硫酸角质素中分离出的多个非还原链末端。

Multiple non-reducing chain termini isolated from bovine corneal keratan sulfates.

作者信息

Tai G H, Huckerby T N, Nieduszynski I A

机构信息

Division of Biological Sciences, Institute of Environmental and Biological Sciences, Lancaster University, Bailrigg, Lancaster LA1 4YQ, United Kingdom.

出版信息

J Biol Chem. 1996 Sep 20;271(38):23535-46. doi: 10.1074/jbc.271.38.23535.

DOI:10.1074/jbc.271.38.23535
PMID:8798563
Abstract

Keratan sulfate-containing proteoglycans were isolated from bovine cornea (15-month-old to 3-year-old animals) and digested with the enzyme, keratanase II. The released oligosaccharides, which included non-reducing termini and repeat region oligosaccharides but not linkage regions, were reduced with alkaline borohydride and fractionated on a Spherisorb column. These oligosaccharides were examined by 600-MHz 1H NMR spectroscopy using one- and two-dimensional methods and, in addition to some oligosaccharide alditols previously recovered from skeletal keratan sulfate, the following new capping structures were identified: NeuAcalpha2-6Galbeta1-4GlcNAc(S)-ol, NeuAcalpha2-3Gal(S)beta1-4GlcNAc(S)beta1-3Galbeta1-4GlcNAc(S )-ol, NeuGcalpha2-6Galbeta1-4GlcNAc(S)beta1-3Galbeta1-4Gl cNA c(S)-ol, NeuGcalpha2-3Galbeta1-4GlcNAc(S)beta1-3Galbeta1-4Gl cNA c(S)-ol, NeuGcalpha2-3Gal(S)beta1-4GlcNAc(S)beta1-3Galbeta1-4GlcNAc(S )-ol, NeuGcalpha2-3Gal(S)beta1-4GlcNAc(S)beta1-3Gal(S)beta1-4GlcNAc(S)-o l, Galalpha1-3Galbeta1-4GlcNAc(S)beta1-3Galbeta1-4GlcNAc( S)-ol, Galalpha1-3Galbeta1-4GlcNAc(S)beta1-3Gal(S)beta1-4GlcNAc(S)- ol, GlcNAc(S)beta1-3Gal(S)beta1-4GlcNAc(S)-ol, and GalNAc(S)beta1-3Gal(S)beta1-4GlcNAc(S)-ol. These structures represent seven families of capping residues, whose relative molar proportions are given in parentheses: NeuAcalpha(2-3)- (12%), NeuAcalpha(2-6)- (41%), NeuGcalpha(2-3)- and NeuGcalpha(2-6)- families (12%), Galalpha(1-3)- (26%), GalNAc(S)beta(1-3)- (5%), and GlcNAc(S)beta(1-3)- (4%). It is not clear, at present, where each of these structures occurs on the bi-antennary N-linked corneal keratan sulfate chains, which themselves occur within three keratan sulfate proteoglycan species. However, examination of the relative proportions of the capping to the repeat structures and knowledge of the average molecular size suggests that the sum of these non-reducing termini represents the caps of two antennae.

摘要

从牛角膜（15个月至3岁的动物）中分离出含硫酸角质素的蛋白聚糖，并用角质酶II进行消化。释放出的寡糖，包括非还原末端和重复区域寡糖，但不包括连接区域寡糖，用碱性硼氢化物还原后在Spherisorb柱上进行分级分离。使用一维和二维方法通过600兆赫兹的1H核磁共振光谱对这些寡糖进行检测，除了之前从骨骼硫酸角质素中回收的一些寡糖醇外，还鉴定出了以下新的封端结构：NeuAcalpha2-6Galbeta1-4GlcNAc(S)-ol、NeuAcalpha2-3Gal(S)beta1-4GlcNAc(S)beta1-3Galbeta1-4GlcNAc(S)-ol、NeuGcalpha2-6Galbeta1-4GlcNAc(S)beta1-3Galbeta1-4GlcNAc(S)-ol、NeuGcalpha2-3Galbeta1-4GlcNAc(S)beta1-3Galbeta1-4GlcNAc(S)-ol、NeuGcalpha2-3Gal(S)beta1-4GlcNAc(S)beta1-3Galbeta1-4GlcNAc(S)-ol、NeuGcalpha2-3Gal(S)beta1-4GlcNAc(S)beta1-3Gal(S)beta1-4GlcNAc(S)-ol、Galalpha1-3Galbeta1-4GlcNAc(S)beta1-3Galbeta…

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