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一种胆碱能神经元分化因子的纯化及部分特性分析

Purification and partial characterization of a cholinergic neuronal differentiation factor.

作者信息

Fukada K

出版信息

Proc Natl Acad Sci U S A. 1985 Dec;82(24):8795-9. doi: 10.1073/pnas.82.24.8795.

DOI:10.1073/pnas.82.24.8795
PMID:3866253
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC391524/
Abstract

The choice of transmitter made by postmitotic rat sympathetic neurons in cell culture can be controlled by the environment in which they develop. One of the differentiation signals is a protein secreted by heart cells that can induce previously noradrenergic neurons to synthesize acetylcholine and form cholinergic synapses. This change in phenotype occurs without alteration in neuronal survival or growth. The differentiation factor has now been purified at least 100,000-fold, and it is homogeneous by several criteria. (i) The cholinergic activity comigrates with a single 125I-labeled protein band of 45 kDa in one-dimensional NaDodSO4/PAGE. (ii) The biological activity comigrates precisely with a series of five 125I-labeled protein spots of 45 kDa in two-dimensional gel electrophoresis. (iii) Treatment of the 45-kDa band with endo-beta-N-acetylglucosaminidase F reduces the apparent molecular size of both the labeled protein and the biological activity to a band of 22 kDa. The data suggest that the differentiation factor is a slightly basic glycoprotein with at least six glycosylation sites.

摘要

细胞培养中，有丝分裂后的大鼠交感神经元对递质的选择可由其发育所处的环境控制。其中一种分化信号是心脏细胞分泌的一种蛋白质，它能诱导先前的去甲肾上腺素能神经元合成乙酰胆碱并形成胆碱能突触。这种表型变化发生时，神经元的存活或生长并未改变。现在，这种分化因子已被纯化了至少10万倍，并且在几个标准下都是均一的。（i）在一维NaDodSO4/PAGE中，胆碱能活性与一条45 kDa的单一125I标记蛋白带共迁移。（ii）在二维凝胶电泳中，生物活性与一系列五条45 kDa的125I标记蛋白斑点精确共迁移。（iii）用内切β-N-乙酰葡糖胺糖苷酶F处理45 kDa条带，会使标记蛋白和生物活性的表观分子大小都降至22 kDa的条带。数据表明，分化因子是一种略带碱性的糖蛋白，至少有六个糖基化位点。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e78c/391524/0b9ed479c803/pnas00364-0534-e.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e78c/391524/44baa3beae10/pnas00364-0533-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e78c/391524/a99a16daf92e/pnas00364-0534-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e78c/391524/ac1231380e8f/pnas00364-0534-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e78c/391524/4d953d05818f/pnas00364-0534-c.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e78c/391524/cb8b7bab7edd/pnas00364-0534-d.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e78c/391524/0b9ed479c803/pnas00364-0534-e.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e78c/391524/44baa3beae10/pnas00364-0533-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e78c/391524/a99a16daf92e/pnas00364-0534-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e78c/391524/ac1231380e8f/pnas00364-0534-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e78c/391524/4d953d05818f/pnas00364-0534-c.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e78c/391524/cb8b7bab7edd/pnas00364-0534-d.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e78c/391524/0b9ed479c803/pnas00364-0534-e.jpg

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