Analysis of the function of human brain carboxypeptidase B and its diagnostic significance in dementia of Alzheimer's type

人脑羧肽酶B的功能分析及其对阿尔茨海默型痴呆的诊断意义

• 批准号: 19591027
• 负责人: MATSUMOTO Akira
• 金额: $ 2.83万
• 依托单位: Himeji Dokkyo University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2007
• 资助国家: 日本
• 起止时间: 2007 至 2009
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-19591027/
• 关键词: アルツマイマー病; 軽度認知機能障害; カルボキシペプチダーゼB; 立体構造認識; モノクロナル抗体; ベータアミロイド; CPB結合蛋白; 結合ポケット; バイマーカー; エピトープ解析; 立体構造; 酵素; 神経科学; 痴呆; トランスレーショナルリサーチ; 脳神経疾患

1. Biochemical and in silico analyses of the CPB-BP binding pocket of human brain carboxypeptidase B (HBCPB) : In an attempt to understand the mechanism of action of HBCPB and its specific binding protein, CPB-BP (1 & 2), we have started an epitope mapping analyses by using antibodies generated based upon primary structures of the both proteins. These monoclonal antibodies (6 clones) exhibit sufficient specificity and sensitivity, and analyses of binding to CPB-BP (1 & 2) and its regulation have enabled classification of these in terms of difference in three-dimensional structures, apart from specificity and sensitivity. This achievement embodies a basis for structural analysis of the binding pocket and SBDD in the next experimental program.2. A search and analysis for CPB-BP and related proteins in human serum. To facilitate HTP analyses for this purpose, the surface plasmon interaction analysis in combination with MALDI- TOF-MS was utilized. Using a peptide ligand corresponding to the C-terminal epitope of CPB, human peripheral serum samples, from which major abundant proteins were removed, were analyzed. In addition to the successful identification molecules previously determined by immunoprecipitation, a new molecule was identified, with which an interaction to the candidate molecule is recognized.3. Immunohistochemical analyses of pathological specimen. All AD brain samples and a portion of brain samples from the normal aged both exhibit CPB-BP2 immunoreactivity in the hippocampus.

1. 人脑羧肽酶B (HBCPB) CPB-BP结合袋的生化和计算机分析：为了了解HBCPB及其特异性结合蛋白CPB-BP（1和2）的作用机制，我们利用基于这两种蛋白初级结构产生的抗体开始了表位定位分析。这些单克隆抗体（6个克隆）表现出足够的特异性和敏感性，通过分析与CPB-BP（1和2）的结合及其调控，除了特异性和敏感性外，还可以根据它们的三维结构差异进行分类。这一成果为下一个实验项目中装订袋和SBDD的结构分析奠定了基础。人血清CPB-BP及其相关蛋白的搜索与分析。为了便于HTP分析，我们将表面等离子体相互作用分析与MALDI- TOF-MS相结合。利用CPB c末端表位对应的肽配体，对主要富集蛋白被去除的人外周血血清样本进行分析。除了先前通过免疫沉淀确定的成功鉴定分子外，还鉴定了一个与候选分子相互作用的新分子。病理标本免疫组化分析。所有AD脑样本和部分正常老年人脑样本在海马区均表现出CPB-BP2免疫反应性。

项目成果

Quantitative Analysis of β-Amyloid Peptides Expressed in Human Cerebrospinal Fluid by an Improved Method of Antibody-Assisted Time-of-Flight Mass Spectrometry

采用改进的抗体辅助飞行时间质谱法定量分析人脑脊液中表达的 β-淀粉样肽

• DOI: 10.1007/s10989-009-9174-3
• 发表时间: 2009
• 期刊: International Journal of Peptide Research and Therapeutics
• 影响因子: 2.5
• 作者: Akira Matsumoto;R. Matsumoto;K. Kadoyama;Taka;S. Matsuyama;O. Midorikawa
• 通讯作者: O. Midorikawa

Quantitative analysis of beta-amyloid peptides expressed in human cerebrospinal fluid by an improved method of antibody-assisted time-of-flight mass spectrometry

通过改进的抗体辅助飞行时间质谱法对人脑脊液中表达的β-淀粉样肽进行定量分析

• 发表时间: 2009
• 期刊: International Journal of Peptide Research and Therapeutics 15
• 作者: Akira Matsumoto;Reiko Matsumoto;Keiichi Kadoyama;Shogo Matsuyama;et al. (6名中1番目)
• 通讯作者: et al. (6名中1番目)

Quantitative analysis of beta-amyloid peptides expressed in human cerebrospinal fluid by an improved method of antibodv-assisted time-of-flight mass spectrometrv.

通过改进的抗体辅助飞行时间质谱法对人脑脊液中表达的β-淀粉样肽进行定量分析。

• 发表时间: 2009
• 期刊: International Journal of Peptide Research and Theraoeutics 15(電子媒体掲載中)
• 作者: Matsumoto A;et al.
• 通讯作者: et al.

Quantiative analysis of beta-amyloid petides expressed in human cerebrospinal fluid by an improved method of antibody-assisted time-of-flight mass spectrometry

通过改进的抗体辅助飞行时间质谱法对人脑脊液中表达的β-淀粉样蛋白肽进行定量分析

• 期刊: Int.J.Peptide Res (in press)
• 作者: Matsumoto;A.;et. al.
• 通讯作者: et. al.

Use of a phosphosensor dye in proteomic analysis of human mutant tau transgenic mouse

磷酸传感器染料在人类突变 tau 转基因小鼠蛋白质组学分析中的应用

• 发表时间: 2009
• 期刊: Neuro-report 20
• 作者: Takano Masaoki;Otani Mieko;Shogo Matsuyama;Akira Matsumoto;et al. (9名中6番目)
• 通讯作者: et al. (9名中6番目)