Biochemical characterization of tau in tauopaty brains

tau蛋白脑中tau蛋白的生化特征

• 批准号: 12680727
• 负责人: HASEGAWA Masato
• 金额: $ 2.56万
• 依托单位: Tokyo Metropolitan Organization for Medical Research (2001-2002)The University of Tokyo (2000)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12680727/
• 关键词: Tau; Phosphorylation; splicing; α-synuclein; Neurodegenrative disease; Cdk5; Cdk5; 脳神経疾患; 痴呆; 老化; プロテオーム; 蛋白質; アルツハイマー病; FTDP-17; タウオパチー; 前頭側頭型痴呆; p35; PHF

(1)Biochemical characterization of tau from tauopathy brainsWe investigated effects of tau gene mutations (Exon10+11, +12, N279K, R5H, L266V) which were identified in Japanese families by analyzing soluble and insoluble tau from the patients, exon trapping assay and in vitro microtubule assembly assay. Hyperphosphorylated 4-repeat (4R) tau depositions were observed in brains of patients with either Exon10+12 or N279. Both R5H and L266V mutations affected the ability to promote microtubule assembly, and tau deposition of both 3R and 4R isoforms were detected in those brains. Analysis of soluble and insoluble tau from patients with Kii ALS-PDC revealed hyperphosphorylated 3R and 4R tau accumulations and normal pattern of soluble tau.(2)Cdk5 activator p25 in AD brainsWe investigated whether Cdk5 activator p25 that is produced by calpain cleavage of p35 is elevated in AD brain. Analysis of 8 AD and 9 control cases have shown that no significant increase of p25 was detected in AD brain. Furthermore, we clearly showed that p35 degraded to p25 during postmortem intervals.(3)Posttranslational modification on α-synucleinWe analyzed α-synuclein deposited in brains with dementia with Lewy body patients using biochemical and proteinchemical techniques, and identified that the insoluble α-synuclein is hyperphosphorylated at Ser129. We also found that a fraction of phosphorylated insoluble α-synuclein is ubiquitinated in α-synucleinopathy lesions, and that amino acid residues 31-109 of α-synuclein constitute Proteinase K resistant core of the filaments

(1)来自tau蛋白病脑部的tau蛋白的生化特征我们通过分析患者的可溶性和不溶性tau蛋白、外显子捕获测定和体外微管测定组装在患者大脑中观察到过度磷酸化的 4 重复 (4R) tau 沉积。 Exon10+12 或 N279 突变都会影响促进微管组装的能力，并且在这些 Kii ALS-PDC 患者的大脑中检测到 3R 和 4R 同种型的 tau 沉积。过度磷酸化的 3R 和 4R tau 积累以及可溶性 tau 的正常模式。(2)Cdk5 激活剂AD 大脑中的 p25 我们研究了由钙蛋白酶裂解 p35 产生的 Cdk5 激活剂 p25 在 AD 大脑中是否升高。对 8 个 AD 和 9 个对照病例的分析表明，在 AD 大脑中没有检测到 p25 的显着增加。 p35 在死后间隔期间降解为 p25。 (3)α-突触核蛋白的翻译后修饰我们分析了沉积在痴呆症患者大脑中的 α-突触核蛋白使用生化和蛋白质化学技术对路易体患者进行研究，并鉴定出不溶性 α-突触核蛋白在 Ser129 位点过度磷酸化，我们还发现一部分磷酸化的不溶性 α-突触核蛋白在 α-突触核蛋白病病变中泛素化，并且氨基酸残基 31-109 被泛素化。 α-突触核蛋白构成丝的蛋白酶 K 抗性核心

项目成果

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Arima K、Kowalska A、Hasekawa M 等人：“两兄弟患有额颞叶痴呆和帕金森病，伴有 tau 基因 N279K 突变”神经学。

Yasuda M, Takamatsu J, D'Souza I, Crowther RA, Kawamata T, Hasegawa M, et al: "A novel mutation at position +12 in the intron following exon 10 of the tau gene in familial frontotemporal dementia (FTD-Kumamoto)"Ann Neurol. 47. 422-429 (2000)

Yasuda M、Takamatsu J、DSouza I、Crowther RA、Kawamata T、Hasekawa M 等人：“家族性额颞叶痴呆 (FTD-Kumamoto) tau 基因外显子 10 后内含子第 12 位的新突变”

Wakabayashi K, Fukushima T, Koide R, Horikawa Y, Hasegawa M, et al: "Juvenile-onset generalized neuroaxonal dystrophy (Hallervorden-Spatz disease) with diffuse neurofibrillary and Lewy body pathology"Acta Neuropathol. 99. 331-336 (2000)

Wakabayashi K、Fukushima T、Koide R、Horikawa Y、Hasekawa M 等人：“青少年发病的全身性神经轴突营养不良（Hallervorden-Spatz 病），伴有弥漫性神经原纤维和路易体病理学”Acta Neuropathol。

Kuzuhara S, Kokubo Y, Sasaki R, Narita Y, Yabana T, Hasegawa M, et al: "Familial amyotrophic lateral sclerosis and parkinsonism-dementia complex of the Kii Peninsula of Japan: clinical and neuropathological study and tau analysis"Ann Neurol. 49. 501-511 (

Kuzuhara S、Kokubo Y、Sasaki R、Narita Y、Yabana T、Hasekawa M 等人：“日本纪伊半岛的家族性肌萎缩侧索硬化症和帕金森痴呆症复合体：临床和神经病理学研究以及 tau 分析”Ann Neurol。

kahle PJ, Neumann M, Hasegawa M, et al.: "Hyperphosphorylation and isolubility of alpha-synuclein in transgenic mouse oligodendrocytes"EMBO Rep. 3. 583-588 (2002)

kahle PJ、Neumann M、Hasekawa M 等人：“转基因小鼠少突胶质细胞中 α-突触核蛋白的过度磷酸化和同溶性”EMBO Rep. 3. 583-588 (2002)