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）分离和表征发育中牙齿的釉质和牙本质基质中的蛋白质。 B）发现导致牙齿疾病的缺陷基因的突变，和 蛋白质组学的方法提出了分离和表征蛋白质正常表达过程中牙齿形成。通过发现新的牙本质和釉质基质蛋白，我们将确定新的候选基因的遗传性牙科疾病的病因，如牙本质/牙釉质发育障碍（AI）和牙本质发育障碍（DGI）。 提出了两个具体目标： SA：1分离和表征发育中的釉质和牙本质的细胞外基质中的分子;和牙本质。 SA 2：鉴定导致釉质形成不稳定和牙本质形成不稳定的基因和突变。 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组分进行了全面的分离和表征。几乎每一种可溶性基质蛋白质的完全解析都是使用二维方法实现的，该方法首先通过等电点分离蛋白质，其次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、N-末端测序、肽图谱和翻译后修饰表征来表征。 在Aim Specific 2中，使用候选基因方法对患有AI或DGI的家族进行遗传研究。常染色体AI的五个候选基因（在4 qll-q21上的釉蛋白和成釉蛋白，在11 q22上的MMP-20，在1 q21 -31上的簇蛋白，和在19ql3.3-ql3.4上的激肽释放酶-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（4 q21上的DSPP和DMP 1），以及在蛋白质组学研究中发现的候选基因将被测试与牙齿疾病的联系。 这项研究将提供基于基因的诊断标准，改善遗传咨询，并为患有遗传性疾病的家庭开发新的预防和治疗策略。