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）Tauopathy brainswe对Tau的生化表征研究了Tau基因突变（Exon10 +11，+12，N279K，R5H，L266V）的影响，这些作用是通过分析患者的固体和溶解性TAU在日本家庭中鉴定出来的，这些患者，Exon捕获测定法和体外捕集微蛋白组合和无限制的小微管。在Exon10+12或N279的患者的大脑中观察到高磷酸化的4重复（4R）TAU沉积。 R5H和L266V突变都会影响促进微管组装的能力，并且在这些大脑中检测到3R和4R同工型的Tau沉积。对KII ALS-PDC患者的固体和不溶性TAU的分析显示，AD Brainswe中CDK5激活剂P25的过度磷酸化的3R和4R TAU的积累和正常模式研究了CDK5激活剂P25是否由P35的Calpain Cleaceage far In Ad Brain升高。对8 AD和9例对照病例的分析表明，在AD大脑中未检测到P25的显着增加。此外，我们清楚地表明，p35在死后间隔期间降解为p25。（3）对α-突触核中的翻译后修饰分析了lewy lewy身体患者使用生物学和蛋白质化学技术的α-突触核蛋白分析的α-突触核蛋白，并鉴定出lewy身体患者的痴呆症，并鉴定出强大的α-核酸 - Ser129。我们还发现，磷酸化的不溶性α-突触核蛋白的一部分在α-突触核疾病病变中被泛素化，并且氨基酸保留了α-突触核蛋白的31-109构成细丝的蛋白酶K抗蛋白酶K的蛋白酶K

项目成果

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。

Arima K, Kowalska A, Hasegawa M, et al: "Two brothers with frontotemporal dementia and parkinsonism with an N279K mutation of the tau gene"Neurology. 54. 1787-1795 (2000)

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, et al.: "Juvenile-onset generalized neuroaxonal dystrophy (Hallervorden-Spats disease) with diffuse neurofibrillary and Lewy body pathology."Acta Neuropathol.. 99. 331-336 (2000)

Wakabayashi,K.、Fukushima,T.、Koide,R.、Horikawa,Y.、Hasekawa 等人：“青少年发病的全身性神经轴索营养不良（Hallervorden-Spats 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。