Analyses on beta-site APP cleaving enzyme (BACE1) of amyloid β protein deposited in brains of patients with Alzheimer's disease

阿尔茨海默病患者大脑中沉积的β淀粉样蛋白的β位APP裂解酶（BACE1）分析

• 批准号: 12670590
• 负责人: TAMAOKA Akira
• 金额: $ 2.43万
• 依托单位: University of Tsukuba
• 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-12670590/
• 关键词: Alzheimer's disease; amyloid β protein; BACE1; APP; アミロイドβ蛋白; BACE-1; BACE; 神経細胞; 星状グリア細胞; βアミロイド前駆体蛋白; ELISA

Alzheimer's disease (AD) is characterized by the extensive deposition of amyloid β protein (Aβ) in brain cortex. Aβ is produced from β-amyloid precursor protein (APP) by β-secretase and γ-secretase. Recently, β-secretase was identified as beta-site APP cleaving enzyme 1 (BACE1). Inhibition against BACE1 activity could decrease Aβ generation, indicating the possibility that antagonistic drugs for BACE1 are therapeutic tools for AD.We produced rabbit polyclonal antibodies against synthetic peptides of BACE1. Using these antibodies (aniti-BACE1 antibodies), BACE1 was characterized in human brains (temporal lobes) by Western blotting and immunohistochemistry.All brain fractions extracted by Tris-saline, 1% Triton X-100 and 0.5% SDS sequentialy were revealed to contain BACE1. In order to compare amounts of BACE1 between AD and control brains, brain samples were directly extracted by 0.5% SDS and analyzed by Western blotting and densitometer. Although the mean level of amounts of BACE1 per m … More g protein in AD brains was significantly decreased, the ratio of BACE1 to MAP2 was significantly increased compared to control brains, indicating that remaining small numbers of neurons in AD brains might generate more amounts of BACE1 than control brain neurons. Digestion of both human brains and recombinant BACE1 by N-glycosidase F altered the molecular weight of BACE1 from 70 to 50kDa, consisting with calculated molecular weight of BACE1 depending on its amino acid residues, suggesting that human brains contained both unmodified BACE1 and its glycosylated form. Immunocytochemical studies employing anti-GFAP and anti-MAP2 antibodies as well as anti-BACE1 antibodies have shown that BACE1 was expressed exclusively in neurons.Taking all these findings together in consideration, remaining neurons after neuronal loss in AD brains might generate more amounts of BACE1 than in controls, indicating that increased activities of BACE1 could be one of the causes of AD, which could justify the development of anti-BACE1 drugs for AD treatment. Less

阿尔茨海默氏病（AD）的特征是淀粉样β蛋白（Aβ）在脑皮质中的广泛沉积。 Aβ是由β-分泌酶和γ-分泌酶从β-淀粉样蛋白前体蛋白（APP）产生的。最近，β-分泌酶被鉴定为β位点应用清洁酶1（BACE1）。对BACE1活性的抑制作用可能会降低Aβ的产生，这表明BACE1的拮抗剂是AD的治疗工具。我们生产了针对BACE1合成肽的兔多克隆抗体。使用这些抗体（ANITI-BACE1抗体），BACE1在人的大脑（颞爱）中通过蛋白质印迹和免疫组织化学进行了表征。所有脑部分数由Tris-saline提取，1％Triton X-100和0.5％SDS序列均显示为包含Bace1。为了比较AD和对照大脑之间的BACE1量，通过0.5％SDS直接提取大脑样品，并通过蛋白质印迹和密度计进行分析。尽管AD大脑中的BACE1的平均Bace1量平均水平显着增加，但与对照大脑相比，BACE1与MAP2的比率显着增加，这表明AD大脑中剩余的少量神经元可能会产生比对照脑神经元更多的BACE1。 N-糖苷酶F对人的大脑和重组BACE1的消化使BACE1的分子量从70升至50KDA，这取决于BACE1的分子量取决于其氨基酸残留物的分子量，这表明人的大脑既包含未经改性的BACE1及其糖基化形式。 Immunocytochemical studies employing anti-GFAP and anti-MAP2 antibodies as well as anti-BACE1 antibodies have Shown that BACE1 was expressed exclusively in neurons.Taking all these findings together in consideration, remaining neurons after neuronal loss in AD brains might generate more amounts of BACE1 than in controls, indicating that increased activities of BACE1 could be one of the causes of AD, which could证明开发抗BACE1药物进行AD治疗。较少的

项目成果

A case of primary focal lingual dystonia induced by speaking

• DOI: 10.1046/j.1468-1331.2001.00276.x
• 发表时间: 2001-09-01
• 期刊: EUROPEAN JOURNAL OF NEUROLOGY
• 影响因子: 5.1
• 作者: Ishii, K;Tamaoka, A;Shoji, S
• 通讯作者: Shoji, S

AIDS脳症にみられた脳トキソプラズマ症

艾滋病脑病中可见脑弓形虫病

• 发表时间: 2000
• 期刊: 神経内科 53・Suppl 2
• 作者: Ishii K;玉岡晃;玉岡晃;玉岡晃;玉岡晃;Ishii K;Ishii K;Watanabe M;吉田秀明;吉田秀明;松下正明;玉岡晃;玉岡晃;古庄健太郎;藤田祐之;Tokuda T;Fujita Y;Miyawaki K;Ishii K;Watanabe M;Matsuno S;Takahiko Tokuda;Fujita Y;Kyoko Miyawaki;玉岡晃;玉岡晃;玉岡晃;玉岡晃;玉岡晃;Tokuda T;Ishii K;星野幸子;河野豊;河野豊;Ishii K;Marsuno S;原田祐嗣;玉岡晃;玉岡晃;Kazuhiro Ishii;Ishii K;Sayoko Matsuno;Takahiko Tokuda;Harada H;Hoshi K;Cavani S;古庄健太郎
• 通讯作者: 古庄健太郎

Pictures in clinical medicine. Fungal endophthalmitis and Churg-Strauss syndrome.

临床医学图片。

• 发表时间: 2002
• 期刊: Intern Med 41(2)
• 作者: Ishii K;玉岡晃;玉岡晃;玉岡晃;玉岡晃;Ishii K;Ishii K;Watanabe M;吉田秀明;吉田秀明;松下正明;玉岡晃;玉岡晃;古庄健太郎;藤田祐之;Tokuda T;Fujita Y;Miyawaki K;Ishii K;Watanabe M;Matsuno S;Takahiko Tokuda;Fujita Y
• 通讯作者: Fujita Y

A case with mysthenia gravis (MG) emerging after splenectomy for idiopathic thrombocytopenic purpura (ITP) : possible effects of thymectomy on autoantibodies.

特发性血小板减少性紫癜（ITP）脾切除后出现重症肌无力（MG）病例：胸腺切除术对自身抗体的可能影响。

• 发表时间: 2001
• 期刊: J Med 31
• 作者: Ishii K;玉岡晃;玉岡晃;玉岡晃;玉岡晃;Ishii K;Ishii K;Watanabe M;吉田秀明;吉田秀明;松下正明;玉岡晃;玉岡晃;古庄健太郎;藤田祐之;Tokuda T;Fujita Y;Miyawaki K;Ishii K;Watanabe M;Matsuno S;Takahiko Tokuda;Fujita Y;Kyoko Miyawaki;玉岡晃;玉岡晃;玉岡晃;玉岡晃;玉岡晃;Tokuda T;Ishii K;星野幸子;河野豊;河野豊;Ishii K;Marsuno S;原田祐嗣;玉岡晃;玉岡晃;Kazuhiro Ishii;Ishii K;Sayoko Matsuno
• 通讯作者: Sayoko Matsuno

Three-dimentional and fractal amalyses of assemblies of amyloid β protein subtypes(AB40 and AB42(93))

β淀粉样蛋白亚型（AB40和AB42（93））组装的三维和分形分析

• 发表时间: 2002
• 期刊: Acta Nauropathol (Berl) 103・3
• 作者: Ishii K;玉岡晃;玉岡晃;玉岡晃;玉岡晃;Ishii K;Ishii K;Watanabe M;吉田秀明;吉田秀明;松下正明;玉岡晃;玉岡晃;古庄健太郎;藤田祐之;Tokuda T;Fujita Y;Miyawaki K
• 通讯作者: Miyawaki K