Developmental Studies in the Skeletal Dysplasias

骨骼发育不良的发育研究

基本信息

批准号：
7245971
负责人：
Brendan Lee
金额：
$ 29.88万
依托单位：
CEDARS-SINAI MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-04-01 至 2012-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7245971
关键词：
2-Oxoglutarate 5-Dioxygenase Procollagen-Lysine 2-oxoglutarate 3-dioxygenase proline Address Affect Amino Acids Anabolism Apoptosis Area Binding Biochemical Biological Assay Biological Process Biology Bone and Cartilage Funding Cartilage Cell Differentiation process Chondrocytes Clinical Collaborations Collagen Collagen Type III Collagen Type IV Data Development Dioxygenases Ehlers-Danlos Syndrome Fibrillar Collagen Gene Family Genes Genetic Goals Human Hydroxylation Hypoxia In Vitro Knockout Mice Kyphosis deformity of spine Modification Mus Mutant Strains Mice Mutation Mutation Analysis Nail-Patella Syndrome Osteoblasts Osteochondrodysplasias Osteogenesis Imperfecta Osteoporosis Pathogenesis Patients Pattern Personal Satisfaction Phenotype Platelet Factor 4 Positioning Attribute Post-Translational Modification Alteration Post-Translational Protein Processing Process Procollagen-Proline Dioxygenase Progress Reports Proline Protein Biochemistry Protein Family Proteins Proteomics Range Recruitment Activity Registries Reporting Research Personnel Signal Pathway Signal Transduction Skeletal Development Specific qualifier value Tissues alpha ketoglutarate human study in vivo loss of function member morphogens novel programs protein protein interaction skeletal dysplasia transcription factor

项目摘要

A poorly understood collagen post-translational modification has been 3-prolyl-hydroxylation (P3H) converting proline to 3-hydroxy proline (3-Hyp). It occurs uniquely in the fibrillar collagens, i.e., types I, II and III collagen, at only one proline position, but abundantly in other collagens such as network collagens, i.e., type IV collagen, where it occurs in up to 10% of residues. This begs the question of whether 3-Hyp residues may serve divergent biological functions in different settings and tissues. In recent data, we have identified a novel protein, CRTAP or Cartilage Associated Protein, that is representative of the Leprecanfamily of proteins recently reported to contain a conserved 2-oxoglutarate dioxygenase domain that is found in collagen 4-prolyl-hydroxylases, Hypoxic Inducible Factor (HIF) 4-prolyl-hydoxylases (PDHs), and lysyl hydroxylases (PLODs). Moreover, Leprecan or P3H1 has collagen 3-prolyl-hydroxylase activity in vitro implicating this family of genes as the long sought after PSH's. By combining human and mouse genetic, and proteomic approaches, we show (see Preliminary Studies C.2.) that loss of Crtap in mice causes an osteochondrodysplasia characterized by short stature, kyphosis, and severe osteoporosis. Moreover, this phenotype is biochemically associated with conversion of the single 3-hydroxy- proline to proline in the triple helical domain of types I and II collagen. CRTAP can bind P3H1 and is required for P3H activity in vivo. Finally, we found that loss of CRTAP is associatedwith recessive osteogenesis imperfecta types VII and ll/lll in humans. These data raise important questions that we will address in our Specific Aims. 1) Are the phenotypic features of Crtap loss of function due solely to loss of 3-prolyl- hydroxylation of fibrillar collagens and are there redundant functions with P3H1? 2) What are the consequences of 3-Hyp loss in cartilage and bone on cellular differentiation and function, and collagen biosynthesis? 3)What is the full human clinical spectrum associated with loss of 3-prolyl-hydroxylation of fibrillar collagens? By combining mouse and human studies, this proposal will interact directly with Projects I, II, and the Protein Biochemistry Core to answer these questions.

经过较少了解的胶原蛋白翻译后修饰是3-羟基羟基化（P3H）将脯氨酸转化为3-羟基脯氨酸（3-HYP）。它是在纤维胶原蛋白的独特发生的，即I型，II和III胶原蛋白，仅在一个脯氨酸位置，但在其他胶原蛋白中大量作为网络胶原蛋白，即IV型胶原蛋白，最多在10％的残基中发生。这乞求 3-HYP残基在不同情况下是否可以发挥不同的生物学功能的问题和组织。在最近的数据中，我们确定了一种新型蛋白质，CRTAP或软骨相关蛋白质，代表着最近报道的蛋白质蛋白质的麻风病。保守的2-氧化甲酸二加氧酶结构域，在胶原蛋白4-丙基 - 羟化酶中发现，低氧诱导因子（HIF）4-丙基 - 羟化酶（PDHS）和莱赛羟基酶（PLODS）。此外，Leprecan或P3H1在体外具有胶原蛋白3-丙基羟化酶活性，这意味着这一家基因家族是长期以来一直在寻求PSH的基因。通过结合人类和小鼠遗传，以及蛋白质组学方法，我们表明（请参阅初步研究C.2。）小鼠CRTAP的丧失会导致骨软骨发育不全，其特征是身材矮小，脑膜炎和严重的骨质疏松症。此外，这种表型在生化上与单个3-羟基的转化有关在I型和II胶原蛋白的三重螺旋结构域中的脯氨酸。 CRTAP可以结合P3H1，并且 P3H活性在体内需要。最后，我们发现CRTAP的损失与人类中隐性成骨的不完美型VII和ll/lll。这些数据提高了重要我们将在具体目标中解决的问题。 1）CRTAP丧失功能的表型特征仅由于3-丙基的损失原纤维胶原的羟基化，P3H1是否有冗余功能？ 2）什么是软骨和骨骼中3-HYP损失对细胞分化和功能和胶原蛋白生物合成？ 3）什么是与人类的完整临床光谱相关的纤维胶原蛋白的3-羟基羟基化丧失？通过组合小鼠和人研究，该建议将直接与项目I，II和蛋白质生物化学核心互动回答这些问题。