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: 6873765
• 负责人: JAMES P SIMMER
• 金额: $ 33.82万
• 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/6873765
• 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)分离和鉴定发育中牙齿的釉质和牙本质基质中的蛋白质。 B)发现导致牙病的缺陷基因的突变，以及 提出了一种蛋白质组学方法来分离和鉴定牙齿形成过程中正常表达的蛋白质。通过发现新的牙本质和釉质基质蛋白，我们将在遗传性牙病的病因中发现新的候选基因，如AME/发育不全(AI)和牙本质发育不全(DGI)。 提出了两个具体目标： SA：1分离和鉴定发育中的牙釉质和牙本质以及牙本质细胞外基质中的分子。 SA 2：识别导致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，我们对发育中的牙釉质和牙本质中的蛋白质成分进行了全面的分离和表征。使用先等电点后疏水分离蛋白质的二维方法，几乎可以完全分辨每一种可溶基质蛋白质。分离出的蛋白质通过在线电喷雾电离飞行时间质谱仪、N-末端测序、肽图谱和翻译后修饰的表征进行表征。 在特定的目的中，使用候选基因方法对患有AI或DGI的家系进行了2项遗传学研究。常染色体AI的五个候选基因(4ql 1-q21上的釉蛋白和成釉蛋白，1lq22上的MMP20，lq21-31上的tuftelin，19ql 3.3-ql 3.4上的kallikrein-4)，X连锁AI的一个候选基因(Xp22.3-p22.1上的釉原蛋白)，DGI的两个候选基因(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用于检测与牙病的连锁。 这项研究将提供基于基因的诊断标准，改进遗传咨询，并导致为患有遗传病的家庭开发新的预防和治疗策略。