Chadalavada D M, Sivakami S
Department of Life Sciences, University of Bombay, India.
Biochem Mol Biol Int. 1997 Aug;42(5):1051-61. doi: 10.1080/15216549700203511.
An acid alpha-glucosidase (EC 3.2.1.3) has been purified to electrophoretic homogeneity from the soluble fraction of the human term placenta. In the presence of SDS, two doublets of 79 and 67 kDa were seen in addition to other bands of extremely low intensity. Each of these bands was seen to cross-react with polyclonal antiserum raised to the purified enzyme, thus confirming the homogeneity of the preparation. The purified enzyme exhibited a broad substrate specificity. The kinetic data revealed the possible presence of multiple substrate binding sites. Chemical modification using group specific reagents indicated the presence of a carboxyl group and tryptophan at the active site. Based on these results a possible structure for the active site of the human term placental acid alpha-glucosidase has been proposed.
已从足月人胎盘的可溶性部分中纯化出一种酸性α-葡萄糖苷酶（EC 3.2.1.3），达到电泳纯。在十二烷基硫酸钠（SDS）存在的情况下，除了其他极低强度的条带外，还观察到79 kDa和67 kDa的两个双峰。这些条带中的每一条都与针对纯化酶产生的多克隆抗血清发生交叉反应，从而证实了制备物的均一性。纯化的酶表现出广泛的底物特异性。动力学数据表明可能存在多个底物结合位点。使用基团特异性试剂进行的化学修饰表明活性位点存在羧基和色氨酸。基于这些结果，提出了足月人胎盘酸性α-葡萄糖苷酶活性位点的可能结构。
