Grabowski G A, Ikonne J U, Desnick R J
Enzyme. 1980;25(1):13-25. doi: 10.1159/000459210.
Human alpha-mannosidases A, B and C were partially purified from liver by conventional and affinity chromatographic procedures. The kinetic, physical and immunologic properties of the A, B and C isozymes were determined including pH optima, Km, effects of various inhibitors and activators, thermal stabilities, electrophoretic mobilities, native molecular weights and antigenic relationships. By gel filtration, the apparent native molecular weights were 240,000, 300,000 and 420,000 for the A, B and C isozymes, respectively. By polyacrylamide gel electrophoresis, the A and B isozymes had native molecular weights of approximately 150,000 and appeared to be charge isomers. Antibodies to the purified A and B isozymes were produced; cross-reactivity between the A and B isozymes against anti-A and anti-B immune sera were observed by double immunodiffusion and immunoprecipitation. In contrast, no reactivity of the C isozyme was detected with either immune serum. Based on these studies, models are presented for the molecular interrelationship of alpha-mannosidases A and B.
通过传统的和亲和层析方法从肝脏中部分纯化了人α-甘露糖苷酶A、B和C。测定了A、B和C同工酶的动力学、物理和免疫学特性，包括最适pH值、Km值、各种抑制剂和激活剂的作用、热稳定性、电泳迁移率、天然分子量和抗原关系。通过凝胶过滤法，A、B和C同工酶的表观天然分子量分别为240,000、300,000和420,000。通过聚丙烯酰胺凝胶电泳，A和B同工酶的天然分子量约为150,000，似乎是电荷异构体。制备了针对纯化的A和B同工酶的抗体；通过双向免疫扩散和免疫沉淀观察到A和B同工酶之间针对抗A和抗B免疫血清的交叉反应性。相比之下，未检测到C同工酶与任何一种免疫血清的反应性。基于这些研究，提出了α-甘露糖苷酶A和B分子相互关系的模型。
