Molecular pathological analysis of citrullinated proteins in the brain from neurodegenerative disorder patients.

神经退行性疾病患者大脑中瓜氨酸蛋白的分子病理学分析。

• 批准号: 14570210
• 负责人: ISHIGAMI Akihito
• 金额: $ 2.11万
• 依托单位: Tokyo Metropolitan Institute of Gerontology
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14570210/
• 关键词: neurodegenerative disorder; Alzheimer's disease; peptidylarginine deiminase; citrullinated protein; myelin basic protein; astrocyte; aging; proteome; 蛋白質脱イミノ化酵素; 脱イミノ化蛋白質; シトルリン; チュブリン; グリア細胞

Citrullinated proteins are the products of a post-translational process in which arginine residues undergo modification into citrulline residues when catalyzed by peptidylarginine deiminases(PADs) in a calcium ion-dependent manner. Enzymatic citrullination abolishes positive charges of native protein molecules, inevitably causing significant alterations in their structure and functions. In mammalian tissues, PADs are found as five different isoforms (i.e, types 1-4,6). PAD2 expressed mainly in the rat cerebrum became activated early in the neurodegenerative process. To elucidate the involvement of the citrullination in neuronal degeneration, we examined whether citrullinated proteins are produced in Alzheimer's disease(AD), Parkinson's disease, and Huntington chorea.1.Histochemical analysis revealed that citrullinated proteins were detected all over the hippocampus. However, no citrullinated proteins were detected in the normal hippocampus. PAD2 immunoreactivity was also detected all over the hippocampus both in the AD and the normal brain. Double immunofluorescence staining revealed that citrullinated protein- and PAD2-positive cells were well-coinddenced with GFAP-positive cells, but not all GFAP-positive cells were PAD2-positive cells. As GFAP is a astrocyte-spedfic marker protein, PAD2 is distributed mainly in astrocytes.2.Citrullinated proteins with various molecular weight were detected in AD hippocampus by Western blot analysis, but not in normal brain. Two of the citrullinated proteins were identified as a vimentin and a glial fibrillary acidic protein(GFAP).Thus, abnormal accumulation of citrullinated proteins and abnormal activation of PAD2 is occurring in AD hippocampus. These results strongly suggested that PAD has an important role in the onset and progression of AD and citrullinated proteins may became a possible useful marker for human neurodegenerative disease.

瓜氨酸蛋白是翻译后过程的产物，其中精氨酸残基在肽基精氨酸脱亚胺酶 (PAD) 以钙离子依赖性方式催化时修饰为瓜氨酸残基。酶促瓜氨酸化消除了天然蛋白质分子的正电荷，不可避免地导致其结构和功能发生显着改变。在哺乳动物组织中，PAD 有五种不同的亚型（即 1-4,6 型）。 PAD2 主要在大鼠大脑中表达，在神经退行性过程的早期被激活。为了阐明瓜氨酸化在神经元变性中的作用，我们检查了阿尔茨海默病（AD）、帕金森病和亨廷顿舞蹈病是否产生瓜氨酸蛋白。1.组织化学分析显示，在整个海马体中都检测到了瓜氨酸蛋白。然而，在正常海马中没有检测到瓜氨酸蛋白。在 AD 和正常大脑的整个海马体中也检测到了 PAD2 免疫反应性。双重免疫荧光染色显示瓜氨酸蛋白和 PAD2 阳性细胞与 GFAP 阳性细胞高度一致，但并非所有 GFAP 阳性细胞都是 PAD2 阳性细胞。由于GFAP是星形胶质细胞特有的标记蛋白，PAD2主要分布在星形胶质细胞中。2.Western blot检测AD海马中检测到不同分子量的瓜氨酸蛋白，而正常脑中未检测到。其中两种瓜氨酸蛋白被鉴定为波形蛋白和神经胶质纤维酸性蛋白(GFAP)。因此，AD海马中出现了瓜氨酸蛋白的异常积累和PAD2的异常激活。这些结果强烈表明，PAD 在 AD 的发生和进展中具有重要作用，瓜氨酸蛋白可能成为人类神经退行性疾病的可能有用标记。

项目成果

Protein deimination and peptidylarginine deiminase expression during cornification of rat epidermal keratinocytes.

大鼠表皮角质形成细胞角质化过程中的蛋白质脱亚胺化和肽基精氨酸脱亚胺酶表达。

• 发表时间: 2002
• 期刊: Biomed.Res. 23
• 作者: Ishigami;A.;Asaga;H.;et al.
• 通讯作者: et al.

Senescence marker protein-30 (SMP30) regulates Akt activity and contributes to cell survival in Hep G2 cells.

衰老标记蛋白 30 (SMP30) 调节 Akt 活性并有助于 Hep G2 细胞的细胞存活。

• 发表时间: 2004
• 期刊: Biochem.Biophys.Res.Commun. 321
• 作者: Matsuyama;S.;Kitamura;T.;Enomoto;N.;Fujita;T.et al.
• 通讯作者: T.et al.

Abnormal accumulation of deiminated proteins catalyzed by peptidylarginine deiminase in hippocampal extracts from patients with Alzheimer's disease.

阿尔茨海默病患者海马提取物中肽基精氨酸脱亚胺酶催化的脱亚胺蛋白异常积累。

• 发表时间: 2005
• 期刊: J.Neurosci.Res. 80
• 作者: Ishigami;A;Ohsawa;T;Hiratsuka;M;Taguchi;H;Kobayashi;S;Saito;Y;Murayama;S;Asaga;H;Toda;T;Kimura;N.Maruyama;N.
• 通讯作者: N.

Mori, T., Ishigami, A.et al.: "Senescence marker protein-30 knockout mouse as a novel murine model of senile lung."Pathol.Int.. 54. 167-173 (2004)

Mori, T., Ishigami, A.等人：“衰老标记蛋白 30 敲除小鼠作为老年肺的新型小鼠模型。”Pathol.Int.. 54. 167-173 (2004)

cDNA cloning, gene organization and expression analysis of human peptidylarginine deiminase type I

• DOI: 10.1042/bj20020870
• 发表时间: 2003-02-15
• 期刊: BIOCHEMICAL JOURNAL
• 影响因子: 4.1
• 作者: Guerrin, M;Ishigami, A;Serre, G
• 通讯作者: Serre, G