Proteomic and Genetics of Enamel and Dentin

牙釉质和牙本质的蛋白质组学和遗传学

• 批准号: 7064910
• 负责人: JAMES P SIMMER
• 金额: $ 28.39万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-04-01 至 2009-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7064910
• 关键词: clinical research; dental development; dental disorder; dentin; developmental genetics; electrospray ionization mass spectrometry; extracellular matrix; extracellular matrix proteins; family genetics; gene mutation; genetic disorder; genetic screening; human subject; isoelectric point; patient oriented research; protein sequence; proteomics; swine; tooth enamel

DESCRIPTION: Proteomics and Genetics of Enamel and Dentin: Our goals are: a) to isolate and characterize proteins in the enamel and dentin matrices of developing teeth. b) to discover mutations in defective genes that cause dental disorders, and A proteomics approach is proposed to isolate and characterize proteins normally expressed during tooth formation. By discovering novel dentin and enamel matrix proteins, we will identify new candidate genes in the etiologies of inherited dental disorders, such as ame/ogenesis imperfecta (AI) and dentinogenesis imperfecta (DGI). Two Specific Aims are proposed: SA:1 Isolate and characterize molecules in the extracellular matrices of developing enamel and dentin; and dentin. SA 2: Identify genes and mutations that cause amelogenesis imperfecta and dentinogenesis imperfecta. In Specific Aim 1 we perform a comprehensive isolation and characterization of the protein components in developing enamel and dentin. Complete resolution of virtually every soluble matrix protein is achieved using a two-dimensional method that separates proteins first by isoelectric point and second by hydrophobicityo Isolated proteins are characterized by on-line electrospray ionization (ESI) time-of-flight mass spectrometry, by N-terminal sequencing, peptide mapping and characterization of posttranslational modifications. In Specific Aim 2 genetic studies are performed on kindreds having AI or DGI, using a candidate gene approach. The five candidate genes for autosomal AI (enamelin & ameloblastin on 4ql 1-q21, MMP-20 on 1lq22, tuftelin on lq21-31, and kallikrein-4 on 19ql 3.3-ql 3.4), the one candidate gene for X-linked AI (amelogenin on Xp22.3-p22.1), the two candidate genes for DGI (DSPP and DMP1 on 4q21), and candidate genes identified discovered in the Proteomics study will be tested for linkage to dental disease. This study will provide gene-based diagnostic criteria, improved genetic counseling, and lead to the development of novel prevention and therapeutic strategies for families suffering from inherited disease.

描述：牙釉质和牙本质的蛋白质组学和遗传学：我们的目标是： a) 分离和表征发育中牙齿的牙釉质和牙本质基质中的蛋白质。 b) 发现导致牙齿疾病的缺陷基因突变，以及 提出了一种蛋白质组学方法来分离和表征牙齿形成过程中通常表达的蛋白质。通过发现新的牙本质和牙釉质基质蛋白，我们将在遗传性牙齿疾病的病因学中识别新的候选基因，例如牙釉质/发育不全症（AI）和牙本质发生不全症（DGI）。 提出了两个具体目标： SA:1 分离并表征发育中的牙釉质和牙本质的细胞外基质中的分子；和牙本质。 SA 2：识别导致牙釉质发育不全和牙本质发育不全的基因和突变。 在具体目标 1 中，我们对牙釉质和牙本质发育过程中的蛋白质成分进行了全面的分离和表征。使用二维方法实现了几乎所有可溶性基质蛋白的完全分辨率，该方法首先通过等电点分离蛋白质，然后通过疏水性分离蛋白质。分离的蛋白质通过在线电喷雾电离 (ESI) 飞行时间质谱法、N -末端测序、肽图谱和翻译后修饰的表征。 在特定目标 2 中，使用候选基因方法对患有 AI 或 DGI 的亲属进行遗传研究。常染色体 AI 的五个候选基因（4ql 1-​​q21 上的牙釉质和成釉细胞、1lq22 上的 MMP-20、lq21-31 上的 tuftelin 和 19ql 3.3-ql 3.4 上的激肽释放酶-4），是 X 连锁 AI 的一个候选基因（Xp22.3-p22.1 上的釉原蛋白），两个候选基因DGI（4q21 上的 DSPP 和 DMP1）以及蛋白质组学研究中发现的候选基因将被测试与牙科疾病的联系。 这项研究将提供基于基因的诊断标准，改进遗传咨询，并为患有遗传性疾病的家庭开发新的预防和治疗策略。