Aletta J M, Angeletti R, Liem R K, Purcell C, Shelanski M L, Greene L A
Department of Pathology, Columbia University College of Physicians and Surgeons, New York, NY 10032.
J Neurochem. 1988 Oct;51(4):1317-20. doi: 10.1111/j.1471-4159.1988.tb03104.x.
Exposure of PC12 cells to nerve growth factor (NGF) has been shown to induce an mRNA that encodes a novel neuronal intermediate filament protein. The findings presented here concern the identity of this filament protein. The major protein in NGF-treated PC12 cell cytoskeletons derived by extraction with 1% Triton X-100 is of apparent Mr = 58,000, focuses by isoelectric focusing as several closely spaced spots of pl 5.6-5.8, and is elevated relative to non-NGF-treated cells. Partial microsequencing of this material reveals 2 internal sequences that are identical to a 14-residue sequence encoded by the NGF-regulated clone 73 mRNA, but not to sequences of other known proteins. An antiserum raised against a 19-residue synthetic peptide corresponding to the deduced C-terminus of the protein encoded by the NGF-regulated clone 73 mRNA specifically recognizes the 58,000-Mr protein. Properties of the 58-kilodalton protein strongly suggest that it corresponds to an intermediate filament protein (peripherin) previously identified in PC12 cells and in peripheral and certain CNS neurons. Identification of the intermediate filament protein encoded by an NGF-induced message should facilitate studies of its regulation and function.
已证明将PC12细胞暴露于神经生长因子(NGF)会诱导一种编码新型神经元中间丝蛋白的mRNA。本文呈现的研究结果涉及这种丝蛋白的特性。用1% Triton X-100提取得到的NGF处理的PC12细胞细胞骨架中的主要蛋白,其表观分子量为58,000,通过等电聚焦聚焦为几个紧密相邻的等电点为5.6 - 5.8的斑点,并且相对于未用NGF处理的细胞有所增加。对该物质进行的部分微测序揭示了2个内部序列,它们与由NGF调节的克隆73 mRNA编码的一个14个残基的序列相同,但与其他已知蛋白质的序列不同。针对与由NGF调节的克隆73 mRNA编码的蛋白质的推导C末端相对应的一个19个残基的合成肽产生的抗血清,能特异性识别58,000分子量的蛋白质。58千道尔顿蛋白的特性强烈表明它对应于先前在PC12细胞以及外周和某些中枢神经系统神经元中鉴定出的一种中间丝蛋白(外周蛋白)。鉴定由NGF诱导的信使编码的中间丝蛋白应有助于对其调节和功能的研究。