Biochemical and cytochemical evidence indicates that coated vesicles in chick embryo myotubes contain newly synthesized acetylcholinesterase.
Biochemical and cytochemical evidence indicates that coated vesicles in chick embryo myotubes contain newly synthesized acetylcholinesterase.
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生化和细胞化学证据表明，鸡胚胎肌管中的涂层囊泡含有新合成的乙酰胆碱酯酶。
DOI：
10.1083/jcb.101.5.1930
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发表时间：
1985-11
期刊：
影响因子：
--
通讯作者：
Fine RE
中科院分区：
文献类型：
--
作者：
Benson RJ;Porter-Jordan K;Buoniconti P;Fine RE
我们从 17 日龄鸡胚骨骼肌中分离出高度纯化的包被囊泡。这些分离的包被囊泡含有潜在的、膜保护形式的乙酰胆碱酯酶(AChE)，如使用Karnovsky和Roots组织化学程序在酶学和形态学上所证明的(J. Histochem. Cytochem.，1964，12：219-221)。通过使用适当的抑制剂，可以显示胆碱酯酶活性对乙酰胆碱具有特异性。还可以得出结论，大多数 AChE 代表可溶性酶，因为它通过重复冻融循环而变得可溶。为了确定包被囊泡相关AChE的起源，我们从培养的鸡胚肌管中分离出包被囊泡，该肌管已经用氟磷酸二异丙酯（一种细胞内和细胞外AChE的基本上不可逆的抑制剂）处理，并且已经允许恢复3小时。这个时间不足以让任何新合成的 AChE 被分泌出来。这些包被的囊泡还主要含有可溶性乙酰胆碱酯酶。这些数据与包被囊泡是新合成的 AChE 细胞内运输的重要中间体的假设相一致。
We have isolated highly purified coated vesicles from 17-d-old chick embryo skeletal muscle. These isolated coated vesicles contain acetylcholinesterase (AChE) in a latent, membrane-protected form as demonstrated enzymatically and morphologically using the Karnovsky and Roots histochemical procedure (J. Histochem. Cytochem., 1964, 12:219- 221). By the use of appropriate inhibitors the cholinesterase activity can be shown to be specific for acetylcholine. It also can be concluded that most of the AChE represents soluble enzyme since it is rendered soluble by repeated freeze-thaw cycles. To determine the origin of the coated vesicle-associated AChE, we have isolated coated vesicles from cultured chick embryo myotubes which have been treated with diisopropylfluorophosphate, an essentially irreversible inhibitor of both intra- and extracellular AChE, and have been allowed to recover for 3 h. This time is not enough to allow any newly synthesized AChE to be secreted. These coated vesicles also contain predominantly soluble AChE. These data are compatible with the hypothesis that coated vesicles are important intermediates in the intracellular transport of newly synthesized AChE.