Proteomics-based approach identifying autoantibodies in Behcet's disease

基于蛋白质组学的方法识别白塞氏病自身抗体

• 批准号: 17590502
• 负责人: TAKEUCHI Toru
• 金额: $ 1.86万
• 依托单位: Osaka Medical College
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2005
• 资助国家: 日本
• 起止时间: 2005 至 2006
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-17590502/
• 关键词: Behecet's disease; autoantibody; proteome; mass spectrometry; electrophoresis; immunoprecipitation; immunoblotting; 二次元電気泳動; Behcet病

We identified autoantigens specific for Behcet's disease by proteomics-based approaches. These approaches were the combination between electrophoresis or immunoprecipitation and mass spectrometry (MS).To detect autoantigens for Behcet's disease, we performed one or two-dimensional electrophoresis using HEp-2 cells and western blotting. There were several spots specific for Behcet's disease, which identified by comparison with sera of normal healthy controls and disease controls.To identify these spots of interest, the gel spots were excised and each piece of the gel spots was reduced, alkylated, and digested with trypsin. The extracted peptides were analyzed with an UltraFlex. The peptide mass fingerprint was used for protein identification from the tryptic fragment size using the Mascot Search engine based on databases. We identified two proteins, α-enolase and annexin-I, which were reported as autoantigens for Behcet's disease. We have also performed the combination between immunoprecipitation and MS for detecting autoantigens. Other proteins, which are candidate autoantigens, were detected by western blotting and immunoprecipitation, and are now under identification by MS.The combination between Immunoblotting or immunoprecipitation and MS is a useful tool for identifying autoantigens for autoimmune diseases such as Behect's disease.

我们通过基于蛋白质组学的方法鉴定了白塞病的特异性自身抗原。这些方法是电泳或免疫沉淀与质谱（MS）的结合。为了检测白塞病的自身抗原，我们使用HEp-2细胞进行单向或双向电泳和Western印迹。通过与正常健康对照和疾病对照的血清比较，有几个白塞病特异性斑点，为了鉴定这些感兴趣的斑点，切下凝胶斑点，并将每片凝胶斑点还原、烷基化并用胰蛋白酶消化。用UltraFlex分析提取的肽。使用基于数据库的Mascot搜索引擎，根据胰蛋白酶片段大小将肽质量指纹用于蛋白质鉴定。我们鉴定了两种蛋白质，α-烯醇化酶和膜联蛋白-I，它们被报道为白塞氏病的自身抗原。我们还进行了免疫沉淀和MS检测自身抗原之间的组合。其他蛋白质，这是候选自身抗原，检测到的蛋白质印迹和免疫沉淀，现在正在鉴定MS。免疫印迹或免疫沉淀和MS之间的组合是一个有用的工具，用于识别自身免疫性疾病，如白塞氏病的自身抗原。

项目成果

Detection and characterization of variant and modified structures of proteins in blood and tissues by mass spectrometry.

通过质谱法检测和表征血液和组织中蛋白质的变异和修饰结构。

• 发表时间: 2006
• 期刊: Mass Spectrom Rev 25
• 作者: Shimizu A;Nakanishi T;et al.
• 通讯作者: et al.

肥大型心筋症を合併した全身性エリテマトーデスの一例

系统性红斑狼疮并发肥厚性心肌病1例

• 发表时间: 2005
• 期刊: 日本臨床免疫学会会誌 28・6
• 作者: Ozono;Ryoji;Junko Koyama;S.Yamaguchi;小谷卓矢
• 通讯作者: 小谷卓矢

Proteomics-based discovery of α-enolase autoantibody in sera of lung adenocarcinoma

基于蛋白质组学的肺腺癌血清中α-烯醇化酶自身抗体的发现

• 发表时间: 2006
• 期刊: Proceedings of 53th ASMS conference on mass spectrometry and allied topics
• 作者: Nakanishi T;Takeuchi T;et al.
• 通讯作者: et al.

Proteomics-based approach identifying autoantibody against peroxiredoxin VI as a novel serum marker in esophageal squamous cell carcinoma

• DOI: 10.1158/1078-0432.ccr-06-1315
• 发表时间: 2006-11-01
• 期刊: CLINICAL CANCER RESEARCH
• 影响因子: 11.5
• 作者: Fujita, Yoshihisa;Nakanishi, Toyofumi;Tanigawa, Nobuhiko
• 通讯作者: Tanigawa, Nobuhiko

Serum protein profile of rheumatoid arthritis treated with anti-TNF therapy (infliximab).

• DOI: 10.1016/j.jchromb.2007.02.028
• 发表时间: 2007-08
• 期刊: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences
• 作者: T. Takeuchi;T. Nakanishi;Yoko Tabushi;Ayu Hata;T. Shoda;T. Kotani;A. Shimizu;T. Takubo;S. Makino;T. Hanafusa