Brunngraber E G, Brown B D
Biochem J. 1967 Apr;103(1):65-72. doi: 10.1042/bj1030065.
Sialomucopolysaccharides were released from the defatted protein residue by the proteolytic action of papain after extraction of rat whole brains with chloroform-methanol (2:1, v/v). Further purification is achieved by dialysis to remove low-molecular-weight fragments and by precipitation of nucleic acids and glucuronic acid-containing mucopolysaccharides by treatment with cetylpyridinium chloride. Gel filtration of the sialomucopolysaccharides through Sephadex G-200 removes the major portion of the impurities that absorb light in the ultraviolet region. The sialomucopolysaccharides were fractionated on DEAE-Sephadex to yield a population of sialomucopolysaccharides that show an increase in N-acetylneuraminic acid content and a decrease in fucose content as the concentration of chloride required to elute the individual components is increased. On gel filtration on Sephadex G-200, those sialomucopolysaccharide molecules rich in N-acetylneuraminic acid and poor in fucose appear to be larger molecules than those rich in fucose and poor in N-acetylneuraminic acid. A structure is proposed in which all sialomucopolysaccharide molecules are assumed to possess the same repeating unit consisting of hexosamine and hexose. The molecules differ from each other in the number of fucose and N-acetylneuraminic acid residues attached to the basic structure. Most of the hexosamine is present as glucosamine, although one fraction was obtained that appeared to contain galactosamine. Most of the hexose present is accounted for as galactose and mannose, although small amounts of glucose were found in some fractions. Methods of analysis for the N-acetylneuraminic acid and hexosamine components of the sialomucopolysaccharides were defined.
用氯仿 - 甲醇(2:1,v/v)提取大鼠全脑后,通过木瓜蛋白酶的蛋白水解作用从脱脂蛋白残渣中释放出唾液酸粘多糖。通过透析去除低分子量片段,并通过用十六烷基吡啶氯化物处理沉淀核酸和含葡萄糖醛酸的粘多糖来实现进一步纯化。唾液酸粘多糖通过Sephadex G - 200进行凝胶过滤,去除了在紫外区域吸收光的大部分杂质。唾液酸粘多糖在DEAE - Sephadex上进行分级分离,得到一组唾液酸粘多糖,随着洗脱各个组分所需氯离子浓度的增加,其N - 乙酰神经氨酸含量增加,岩藻糖含量降低。在Sephadex G - 200上进行凝胶过滤时,富含N - 乙酰神经氨酸且岩藻糖含量低的那些唾液酸粘多糖分子似乎比富含岩藻糖且N - 乙酰神经氨酸含量低的分子更大。提出了一种结构,其中假设所有唾液酸粘多糖分子都具有由己糖胺和己糖组成的相同重复单元。这些分子彼此的区别在于连接到基本结构上的岩藻糖和N - 乙酰神经氨酸残基的数量。大部分己糖胺以葡萄糖胺形式存在,尽管获得了一个似乎含有半乳糖胺的组分。存在的大部分己糖为半乳糖和甘露糖,尽管在一些组分中发现了少量葡萄糖。定义了唾液酸粘多糖中N - 乙酰神经氨酸和己糖胺成分的分析方法。