Rotundo R L
Proc Natl Acad Sci U S A. 1984 Jan;81(2):479-83. doi: 10.1073/pnas.81.2.479.
The synthesis, assembly, and processing of the multiple molecular forms of acetylcholinesterase (AcChoEase; acetylcholine acetylhydrolase, EC 3.1.1.7) in quail muscle cultures was studied by using lectins to distinguish enzyme molecules residing in different subcellular compartments. Special emphasis was given to the assembly of asymmetric AcChoEase molecules because these appear to be the predominant, if not unique, forms of AcChoEase at the vertebrate neuromuscular junction. All cell surface and secreted AcChoEase forms bind to immobilized wheat germ agglutinin, ricin, and concanavalin A, indicating that they have complex oligosaccharides. After treatment of muscle cells with a membrane-permeable irreversible AcChoEase inhibitor, there is a rapid reappearance of the globular monomeric, dimeric, and tetrameric AcChoEase forms. However, the collagen-tailed asymmetric form does not appear until about 90 min after treatment. Analysis of the AcChoEase oligosaccharides with lectins indicates maturation to complex forms over a 90-min period. A large fraction of the intracellular globular AcChoEase molecules bind only to concanavalin A, indicating that they are assembled in the rough endoplasmic reticulum. In contrast, all intracellular asymmetric AcChoEase binds to wheat germ agglutinin, and a significant fraction binds to ricin, indicating that this unique AcChoEase form is assembled from subunits that have previously acquired complex sugars. I conclude that assembly of asymmetric AcChoEase, hence acquisition of information specifying basal lamina localization, occurs in the Golgi apparatus.
通过使用凝集素来区分存在于不同亚细胞区室中的酶分子,研究了鹌鹑肌肉培养物中乙酰胆碱酯酶(AcChoEase;乙酰胆碱乙酰水解酶,EC 3.1.1.7)多种分子形式的合成、组装和加工。特别强调了不对称AcChoEase分子的组装,因为这些分子似乎是脊椎动物神经肌肉接头处AcChoEase的主要形式(如果不是唯一形式的话)。所有细胞表面和分泌的AcChoEase形式都能与固定化的麦胚凝集素、蓖麻毒素和伴刀豆球蛋白A结合,这表明它们具有复杂的寡糖。在用膜可渗透的不可逆AcChoEase抑制剂处理肌肉细胞后,球状单体、二聚体和四聚体AcChoEase形式迅速重新出现。然而,胶原尾不对称形式直到处理后约90分钟才出现。用凝集素分析AcChoEase寡糖表明,在90分钟内成熟为复杂形式。大部分细胞内球状AcChoEase分子仅与伴刀豆球蛋白A结合,这表明它们在内质网中组装。相比之下,所有细胞内不对称AcChoEase都与麦胚凝集素结合,并且相当一部分与蓖麻毒素结合,这表明这种独特的AcChoEase形式是由先前已获得复杂糖的亚基组装而成。我的结论是,不对称AcChoEase的组装,即获得指定基底层定位的信息,发生在高尔基体中。