Identification of an Axotomy-Induced Glycosylated Protein, AIGP1, Possibly Involved in Cell Death Triggered by Endoplasmic Reticulum–Golgi Stress
Identification of an Axotomy-Induced Glycosylated Protein, AIGP1, Possibly Involved in Cell Death Triggered by Endoplasmic Reticulum–Golgi Stress
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DOI：
10.1523/jneurosci.22-24-10751.2002
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发表时间：
2002-12
期刊：
影响因子：
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通讯作者：
S. Aoki;Q. Su;Hang Li;K. Nishikawa;K. Ayukawa;Y. Hara;K. Namikawa;S. Kiryu-Seo;H. Kiyama;K. Wada
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文献类型：
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作者：
S. Aoki;Q. Su;Hang Li;K. Nishikawa;K. Ayukawa;Y. Hara;K. Namikawa;S. Kiryu-Seo;H. Kiyama;K. Wada
我们开发了一种新的方法，称为n -连锁糖基化信号（NGS）差异显示(DD)-PCR，该方法能够识别编码n -连锁糖基化分子的基因，这些分子表现出不同的表达模式。利用这一创新技术，我们发现了一个n -链糖基化的11-跨膜结构域蛋白，该蛋白在轴切反应中上调。表达量在切开后第3天升高，约在术后第5-7天达到最高水平，随后在第20天逐渐下降。该蛋白被命名为axotomy-induced glycosylated/Golgi-complex protein 1 （AIGP1）。AIGP1的免疫反应性特异性地定位于神经元，并在高尔基体中具有亚细胞定位，表明AIGP1是一种常驻高尔基蛋白。此外，培养神经元中的AIGP1基因表达可被内质网(ER) -高尔基应激源tunicamycin和brefeldin a特异性诱导。我们观察到，AIGP1过表达会增加细胞死亡的频率，并且与活性相关的相应蛋白区域涉及大的第8和第9跨膜环。我们的研究结果表明，AIGP1基因的激活和高尔基体复合体中蛋白质的积累可能有助于受损神经元程序性细胞死亡过程中的信号传导。
We developed a new method, designated N-linked glycosylation signal (NGS) differential display (DD)-PCR, that enables the identification of genes encoding N-linked glycosylated molecules that exhibit varying patterns of expression. Using this innovative technique, we identified an N-linked glycosylated 11-transmembrane domain protein that is upregulated in response to axotomy. Expression levels increased 3 d after axotomy, reached maximal levels at approximately postoperative days 5–7, and then gradually decreased through day 20. The protein was termed axotomy-induced glycosylated/Golgi-complex protein 1 (AIGP1). AIGP1 immunoreactivity is specifically localized in neurons, with subcellular localization within the Golgi, indicating that AIGP1 is a resident Golgi protein. Moreover, AIGP1 gene expression in cultured neurons is specifically induced by the endoplasmic reticulum (ER)–Golgi stressors tunicamycin and brefeldin A. We observed that the frequency of cell death is increased by AIGP1 overexpression and that the corresponding region of the protein implicated in the activity involves the large eighth and ninth transmembrane loops. Our results suggest that AIGP1 gene activation and protein accumulation in the Golgi complex in response to axotomy-induced ER–Golgi stress may contribute to signaling during programmed cell death in damaged neurons.