Keller K L, Keller J M, Moy J N
Biochemistry. 1980 May 27;19(11):2529-36. doi: 10.1021/bi00552a035.
A difference in the extent of sulfation between the heparan sulfate isolated from Swiss 3T3 mouse cells and that from Swiss 3T3 cells transformed by the DNA virus SV40 has been reported previously. This variance is manifested by different chromatographic and electrophoretic properties. Heparan sulfates from the two cell types were treated with nitrous acid under conditions that gave selective deaminative cleavage of glucosaminyl residues with sulfated amino groups in order to define the nature of the difference in sulfation further. The O-sulfate containing fragments from the heparan sulfates were compared by gel filtration and ion-exchange chromatography. The results showed that the 3T3 heparan sulfate contains 8% more O-sulfate than does the SV3T3 heparan sulfate. Analysis of uronic acids revealed that both types of heparan sulfates contain 45% L-iduronic acid and 55% D-glucuronic acid. These and other observations indicate that the primary difference in sulfation between the 3T3 and SV3T3 heparan sulfates lies in the extent of O-sulfation.
先前已有报道称,从瑞士3T3小鼠细胞中分离出的硫酸乙酰肝素与由DNA病毒SV40转化的瑞士3T3细胞中分离出的硫酸乙酰肝素在硫酸化程度上存在差异。这种差异表现为不同的色谱和电泳特性。为了进一步确定硫酸化差异的性质,在能选择性地对带有硫酸化氨基的葡糖胺残基进行脱氨基裂解的条件下,用亚硝酸处理了这两种细胞类型的硫酸乙酰肝素。通过凝胶过滤和离子交换色谱对硫酸乙酰肝素中含O-硫酸盐的片段进行了比较。结果表明,3T3硫酸乙酰肝素的O-硫酸盐含量比SV3T3硫酸乙酰肝素多8%。糖醛酸分析表明,两种类型的硫酸乙酰肝素均含有45%的L-艾杜糖醛酸和55%的D-葡糖醛酸。这些以及其他观察结果表明,3T3和SV3T3硫酸乙酰肝素在硫酸化方面的主要差异在于O-硫酸化的程度。