Dysregulation of 3-prolyl-hydroxylation in Human Skeletal Dysplasias

人类骨骼发育不良中 3-脯氨酰羟基化的失调

• 批准号: 7252537
• 负责人: Brendan Lee
• 金额: $ 11.54万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-08-01 至 2011-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7252537
• 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; Collagen; Collagen Type III; Collagen Type IV; Complex; Data; Dioxygenases; Ehlers-Danlos Syndrome; Event; 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; 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; Proline; Protein Family; Proteins; Proteomics; Range; Recruitment Activity; Registries; Reporting; Signal Pathway; Signal Transduction; Specific qualifier value; Tissues; alpha ketoglutarate; in vivo; loss of function; morphogens; novel; protein protein interaction; skeletal dysplasia

DESCRIPTION (provided by applicant): A poorly understood collagen post-translational modification has been 3-prolyl-hydroxylation (P3H) converting proline to 3-hydroxy proline (3-Hyp). It occurs in the fibrillar collagens, i.e., types I, II and III collagen, at only 1 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 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 Leprecan family of proteins recently reported to contain a conserved 2-oxoglutarate dioxygenase domain that is found in collagen 4-prolyl-hydroxylases (P4Hs), 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.1.) 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. This data raises important mechanistic 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? 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 human clinical spectrum associated with loss of 3-prolyl-hydroxylation of fibrillar collagens? 4) What regulates the context- dependent function of CRTAP? These questions address what we believe to be a new area in matrix biology and pathogenesis of skeletal dysplasias, i.e., the in vivo phenotypic and biochemical consequences of dysregulation of the 3-prolyl-hydroxylation machinery.

描述（由申请人提供）：经过熟悉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）。它发生在原纤维胶原蛋白中，即I型，II和III胶原蛋白，仅在1个脯氨酸位置，但在其他胶原蛋白（例如网络胶原蛋白）中，即IV型胶原蛋白型，最多在10％的残基中发生。这就提出了一个问题，即3-HYP残基是否在不同的环境和组织中发挥不同的生物学功能。 In recent data, we have identified a novel protein, CRTAP or Cartilage Associated Protein, that is representative of the Leprecan family of proteins recently reported to contain a conserved 2-oxoglutarate dioxygenase domain that is found in collagen 4-prolyl-hydroxylases (P4Hs), Hypoxic Inducible Factor (HIF) 4-prolyl-hydoxylases (PDHs), and赖氨酸羟化酶（PLODS）。此外，Leprecan或P3H1在体外具有胶原蛋白3-丙基 - 羟化酶活性，这意味着该基因家族是PSH的长期以来所追求的。通过结合人类和小鼠的遗传和蛋白质组学方法，我们表明（参见初步研究C.1。）小鼠CRTAP的丧失会导致骨软骨发育不良，其特征是身材矮小，脑杆菌和严重的骨质疏松症。此外，这种表型在生化上与I型和II型胶原蛋白的三螺旋结构域中的单个3-羟基丙酸酯转化为脯氨酸。 CRTAP可以结合P3H1，并且在体内P3H活性所需。这些数据提出了我们将在具体目标中解决的重要机理问题。 1）CRTAP丧失功能的表型特征是否仅由于纤维胶原的3-羟基羟基化的损失？ 2）软骨和骨骼中3-HYP损失对细胞分化和功能以及胶原蛋白生物合成的后果是什么？ 3）人类的临床光谱与纤维胶原蛋白的3-羟基羟基化的丧失有关？ 4）是什么调节CRTAP的上下文依赖性功能？这些问题介绍了我们认为是基质生物学和骨骼发育异常发病机理的新领域，即3-丙基 - 羟基化机制的体内表型和生化后果。