猪肾的一种三唾液酸神经节苷脂GT3。体外结构与生物合成。
A trisialosyl ganglioside GT3 of hog kidney. Structure and biosynthesis in vitro.
作者信息
Murakami-Murofushi K, Tadano K, Koyama I, Ishizuka I
出版信息
J Biochem. 1981 Dec;90(6):1817-20. doi: 10.1093/oxfordjournals.jbchem.a133660.
A trisialosyl ganglioside was isolated from hog kidney cortex and purified with a yield of 6.8 nmol/g tissue or 13 mol % of total gangliosides. The homogeneous ganglioside was subjected to graded neuraminidase treatment, mild acid hydrolysis, periodate oxidation-borohydride reduction, permethylation analysis and chromium trioxide oxidation. The results suggested that the structure of this ganglioside is NeuAc alpha 2-8NeuAc alpha 2-8NeuAc alpha 2-3Gal beta 1-4Glc beta 1-1Cer (GT3). On the addition of exogenous disialosyl ganglioside GD3 and radioactive CMP-NeuAc, a GT3 synthesis activity was demonstrated in a crude membrane fraction of hog kidney. The activity was greatest in the presence of detergents and had a pH optimum of 6.4. Apparent Km values for CMP-NeuAc and GD3 were about 0. 6 and 0.1 mM, respectively. The reaction produce was assigned as GT3 by chromatographic separation, carbohydrate analysis and periodate oxidation followed by borohydride reduction.
从猪肾皮质中分离出一种三唾液酸神经节苷脂，并进行了纯化，产量为6.8 nmol/g组织或占总神经节苷脂的13 mol%。将该均一的神经节苷脂进行分级神经氨酸酶处理、温和酸水解、高碘酸盐氧化-硼氢化物还原、全甲基化分析和三氧化铬氧化。结果表明，该神经节苷脂的结构为NeuAcα2-8NeuAcα2-8NeuAcα2-3Galβ1-4Glcβ1-1Cer（GT3）。添加外源性二唾液酸神经节苷脂GD3和放射性CMP-NeuAc后，在猪肾粗膜组分中显示出GT3合成活性。该活性在有去污剂存在时最大，最适pH为6.4。CMP-NeuAc和GD3的表观Km值分别约为0.6和0.1 mM。通过色谱分离、碳水化合物分析以及高碘酸盐氧化后硼氢化物还原，将反应产物鉴定为GT3。
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