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.

瓜氨酸化蛋白是翻译后过程的产物，当肽精氨酸脱亚胺酶（pad）以钙离子依赖的方式催化时，精氨酸残基被修饰成瓜氨酸残基。酶法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的重大改变。在哺乳动物组织中，pad有五种不同的亚型（即1-4,6型）。主要在大鼠大脑中表达的PAD2在神经退行性过程中较早被激活。为了阐明瓜氨酸化在神经元变性中的作用，我们研究了瓜氨酸化蛋白是否在阿尔茨海默病（AD）、帕金森病和亨廷顿舞蹈病中产生。组织化学分析显示，瓜氨酸化蛋白遍布海马。然而，在正常海马中未检测到瓜氨酸化蛋白。在阿尔茨海默病和正常大脑的海马区也检测到PAD2免疫反应性。双免疫荧光染色显示瓜氨酸化蛋白和pad2阳性细胞与gfap阳性细胞一致，但并非所有gfap阳性细胞都是pad2阳性细胞。由于GFAP是星形细胞特异性标记蛋白，PAD2主要分布在星形细胞中。Western blot分析在AD海马中检测到不同分子量的瓜氨酸化蛋白，而在正常脑中未检测到。两个瓜氨酸化蛋白被鉴定为波形蛋白和胶质纤维酸性蛋白（GFAP）。因此，AD海马中出现瓜氨酸蛋白的异常积累和PAD2的异常激活。这些结果强烈提示PAD在AD的发生和发展中起重要作用，瓜氨酸化蛋白可能成为人类神经退行性疾病的有用标记物。

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.

Modulation of gene expression of SMP-30 by LPS and calorie restriction during aging process

• DOI: 10.1016/j.exger.2004.04.005
• 发表时间: 2004-08-01
• 期刊: EXPERIMENTAL GERONTOLOGY
• 影响因子: 3.9
• 作者: Jung, KJ;Ishigami, A;Chung, HY
• 通讯作者: Chung, HY

SMP30 deficiency in mice causes an accumulation of neutral lipids and phospholipids in the liver and shortens the life span

• DOI: 10.1016/j.bbrc.2004.01.091
• 发表时间: 2004-03-12
• 期刊: BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
• 影响因子: 3.1
• 作者: Ishigami, A;Kondo, Y;Maruyama, N
• 通讯作者: Maruyama, N

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.

Human peptidylarginine deiminase type II: molecular cloning, gene organization, and expression in human skin

• DOI: 10.1016/s0003-9861(02)00516-7
• 发表时间: 2002-11-01
• 期刊: ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS
• 影响因子: 3.9
• 作者: Ishigami, A;Ohsawa, T;Maruyama, N
• 通讯作者: Maruyama, N