Induction of Peptidylarginine Deiminase 2 and 3 by Dibutyryl cAMP via cAMP-PKA Signaling in Human Astrocytoma U-251MG cells
Induction of Peptidylarginine Deiminase 2 and 3 by Dibutyryl cAMP via cAMP-PKA Signaling in Human Astrocytoma U-251MG cells
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DOI：
10.1002/jnr.23959
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发表时间：
2017-07-01
影响因子：
4.2
通讯作者：
Ishigami, Akihito
中科院分区：
文献类型：
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作者：
Masutomi, Hirofumi;Kawashima, Saki;Ishigami, Akihito
肽基精氨酸脱亚胺酶（PAD）是一种翻译后修饰酶，以钙依赖性方式将蛋白质精氨酸残基瓜氨酸化（脱亚胺化），产生瓜氨酸残基。酶促瓜氨酸消除天然蛋白质分子的正电荷，不可避免地导致其结构和功能的显著改变。以前，我们报道了瓜氨酸化蛋白的异常积累和阿尔茨海默病患者的大脑中PAD 2含量的增加。在这项研究中，我们研究了PAD的表达，通过使用二丁酰cAMP（dbcAMP）在人星形细胞瘤U-251 MG细胞。在正常培养条件下，用定量PCR检测U-251 MG细胞中PAD 2和PAD 3 mRNA的表达。以剂量依赖性方式添加dbcAMP显著增加了该mRNA表达和蛋白水平。此外，PAD酶活性也显著增加，并呈剂量依赖性。cAMP依赖性PKA抑制剂KT 5720可抑制PAD 2和PAD 3 mRNA的表达，提示U-251 MG细胞中dbcAMP诱导的PAD 2和PAD 3 mRNA的表达是通过cAMP-PKA信号通路介导的。这是第一份报告，以文件的PAD 2和PAD 3 mRNA的表达诱导dbcAMP，并归因于这些基因的诱导介导的cAMP-PKA信号通路在U-251 MG细胞。(C)2016 Wiley Periodicals，Inc.
Peptidylarginine deiminases (PADs) are posttranslational modification enzymes that citrullinate (deiminate) protein arginine residues in a calcium-dependent manner, yielding citrulline residues. Enzymatic citrullination abolishes positive charges of native protein molecules, inevitably causing significant alterations in their structure and function. Previously, we reported the abnormal accumulation of citrullinated proteins and an increase of PAD2 content in hippocampi of patients with Alzheimer disease. In this study, we investigated PAD expression by using dibutyryl cAMP (dbcAMP) in human astrocytoma U-251MG cells. Under normal culture conditions, PAD2 and PAD3 mRNA expression is detectable with quantitative PCR in U-251MG cells. The addition of dbcAMP in a dose-dependent manner significantly increased this mRNA expression and protein levels. Moreover, PAD enzyme activity also increased significantly and dose-dependently. Furthermore, the expression of PAD2 and PAD3 mRNA was inhibited by the cAMP-dependent PKA inhibitor KT5720, suggesting that such expression of dbcAMP-induced PAD2 and PAD3 mRNA is mediated by the cAMP-PKA signaling pathway in U-251MG cells. This is the first report to document the PAD2 and PAD3 mRNA expression induced by dbcAMP and to attribute the induction of these genes to mediation by the cAMP-PKA signaling pathway in U-251MG cells. (C) 2016 Wiley Periodicals, Inc.