Cawthon R M, Pintar J E, Haseltine F P, Breakefield X O
J Neurochem. 1981 Aug;37(2):363-72. doi: 10.1111/j.1471-4159.1981.tb00464.x.
[3H]Pargyline-labeled polypeptides associated with the A and B types of monoamine oxidase (MAO) activity in human tissues were analyzed by sodium dodecyl sulfate--polyacrylamide gel electrophoresis (SDS-PAGE). [3H]Pargyline was bound to MAO A in a crude mitochondrial fraction from the placental trophoblast of a male newborn and to MAO B in blood platelets from the umbilical vein of the same newborn. [3H]Pargyline was also bound to MAO A and B in a crude mitochondrial fraction from cultured skin fibroblasts of a male adult and to MAO B in blood platelets from the same individual. Specific labeling of proteins associated with type A or type B activity in fibroblast cells was achieved by preincubation with selective B or A inhibitors, respectively. For all tissues, SDS-PAGE of [3H]pargyline-bound samples revealed a labeled protein band of apparent molecular weight 63,000 for MAO A and 60,000 for MAO B. When SDS-solubilized, [3H]pargyline-labeled MAO A and B proteins from the same male newborn were subjected to limited proteolysis and one-dimensional peptide mapping in SDS gels, different patterns of [3H]pargyline-labeled peptides were obtained. These findings indicate that distinct enzyme molecules are associated with the A and B types of human MAO activity.
采用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）分析了与人体组织中A、B两种类型单胺氧化酶（MAO）活性相关的[3H]帕吉林标记多肽。[3H]帕吉林与一名男性新生儿胎盘滋养层粗线粒体部分中的MAO A结合，与同一新生儿脐静脉血血小板中的MAO B结合。[3H]帕吉林还与一名成年男性培养皮肤成纤维细胞的粗线粒体部分中的MAO A和B结合，以及与同一个体血血小板中的MAO B结合。分别通过与选择性B或A抑制剂预孵育，实现了对成纤维细胞中与A或B型活性相关蛋白质的特异性标记。对于所有组织，[3H]帕吉林结合样品的SDS-PAGE显示，MAO A的标记蛋白带表观分子量为63000，MAO B为60000。当对来自同一男性新生儿的经SDS溶解的[3H]帕吉林标记的MAO A和B蛋白进行有限蛋白酶解和SDS凝胶中的一维肽图谱分析时，获得了不同模式的[3H]帕吉林标记肽。这些发现表明，不同的酶分子与人类MAO的A、B两种类型活性相关。
