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

阿尔茨海默病（AD）的特征是β淀粉样蛋白（Aβ）在大脑皮层中广泛沉积。Aβ是由β-淀粉样前体蛋白（APP）通过β-分泌酶和γ-分泌酶产生的。作为 β 位点 APP 裂解酶 1 (BACE1)，抑制 BACE1 活性可以减少 Aβ 的产生，表明 BACE1 拮抗药物可能是 AD 的治疗工具。针对 BACE1 合成肽的多克隆抗体。使用这些抗体（抗 BACE1 抗体），通过蛋白质印迹和免疫组织化学对人脑（颞叶）中的 BACE1 进行表征。所有脑组分均通过 Tris-saline、1% Triton X-100 和0.5% SDS 连续被发现含有 BACE1 为了比较 AD 和对照大脑之间的 BACE1 含量，直接采集大脑样本。通过 0.5% SDS 提取并通过蛋白质印迹和光密度计进行分析，尽管 AD 大脑中每 m g 蛋白的平均水平显着降低，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比对照大脑神经元产生更多量的 BACE1。N-糖苷酶 F 对人脑和重组 BACE1 的消化改变了分子量。 BACE1 的分子量从 70 到 50kDa，由根据其氨基酸残基计算出的 BACE1 分子量组成，表明人脑中含有未修饰的 BACE1 及其糖基化形式，使用抗 GFAP 和抗 MAP2 抗体以及抗 GFAP 和抗 MAP2 抗体进行免疫细胞化学研究。 BACE1 抗体表明 BACE1 在神经元中表达。将所有这些发现综合考虑，AD 大脑中神经元损失后剩余的神经元可能会产生更多数量的BACE1 活性高于对照组，表明 BACE1 活性增加可能是 AD 的原因之一，这可以证明开发抗 BACE1 药物用于 AD 治疗是合理的。

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