Romeo G, Di Matteo G, D'urso M, Li S C, Li Y T
Biochim Biophys Acta. 1975 Jun 24;391(2):349-60. doi: 10.1016/0005-2744(75)90259-4.
It has been previously reported that following neuraminidase treatment alpha-galactosidase A is converted into the B form, as revealed by electrophoresis. By a variety of techniques such as isoelectrofocusing, DEAE-chromatography and by enzyme kinetic parameters, no conversion of alpha-galactosidase A into B, or the reverse, could be detected after neuraminidase treatment. Only an apparent transformation of alpha-galactosidase A into B was revealed by Cellogel electrophoresis. In addition, a discrepancy was noticed between the pattern of electrophoretic migration on starch gel and Cellogel and the net electrical charges of the two alpha-galactosidases as deduced by isoelectrofocusing and DEAE-cellulose. Neuraminidase treatment did not affect the activity of alpha-galactosidase A towards the natural substrate, ceramidetrihexoside, but the activity of alpha-galactosidase B decreased by about 30% under the same conditions. The two forms of alpha-galactosidases A and B used in this study were extensively purified by classical procedures.
先前有报道称，经神经氨酸酶处理后，α-半乳糖苷酶A会转化为B型，这一点通过电泳得以揭示。然而，运用等电聚焦、DEAE色谱等多种技术以及酶动力学参数分析，在神经氨酸酶处理后，未检测到α-半乳糖苷酶A向B型的转化，反之亦然。仅通过Cellogel电泳显示出α-半乳糖苷酶A有明显向B型的转变。此外，淀粉凝胶和Cellogel上的电泳迁移模式与通过等电聚焦和DEAE-纤维素推导得出的两种α-半乳糖苷酶的净电荷之间存在差异。神经氨酸酶处理并不影响α-半乳糖苷酶A对天然底物神经酰胺三己糖苷的活性，但在相同条件下，α-半乳糖苷酶B的活性降低了约30%。本研究中使用的两种形式的α-半乳糖苷酶A和B通过经典方法进行了广泛纯化。
