Proteomics of ciliopathy protein complexes

纤毛病蛋白复合物的蛋白质组学

• 批准号: 9470242
• 负责人: Claire D McWhite
• 金额: $ 3.46万
• 依托单位: UNIVERSITY OF TEXAS AT AUSTIN
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-01-15 至 2020-01-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9470242
• 关键词: Animals; Biochemical; Biological; Biological Models; Biological Process; Biology; Candidate Disease Gene; Cilia; Clustered Regularly Interspaced Short Palindromic Repeats; Collection; Comparative Biology; Complex; Congenital Abnormality; Core Protein; Data; Data Set; Databases; Defect; Development; Disease; Embryo; Epithelium; Event; Eye; Fractionation; Future; Genes; Genetic screening method; Hand; Health; Hereditary Disease; Human; Human Genetics; Investigation; Joubert syndrome; Kidney; Knock-out; Knockout Mice; Link; Machine Learning; Maps; Mass Spectrum Analysis; Measures; Methods; Modeling; Molecular Biology; Mus; Mutation; Noise; Organism; Outcome; Pattern; Phenotype; Plant Proteins; Plants; Process; Protein-Protein Interaction Map; Proteins; Proteomics; Research; Resources; Route; Sampling; Series; Signal Transduction; Syndrome; System; Techniques; Testing; Whole Organism; Xenopus; Xenopus laevis; base; biological systems; ciliopathy; cilium biogenesis; comparative; exome; experimental study; heart function; in vivo; link protein; mental function; novel; protein complex; protein protein interaction; reproductive function; skeletal

Summary Ciliopathies are a diverse class of developmental diseases that can manifest as defects in kidney, skeletal, eye, heart, reproductive, or mental function. Many ciliopathies, including Bardet-Biedl, Joubert, and Meckel Gruber Syndromes, are caused by defects in the major protein complexes that characterize the ciliary compartment. At least 1,000 proteins are associated with cilia biogenesis and function; uncovering the principles of ciliary protein organization thus requires a large scale, systematic investigation. Our current human protein complex maps, while extensive, have only moderate coverage of ciliary proteins, warranting the collection of targeted experimental datasets. I will use comparative proteomic analysis of cilia from multiple organisms to determine deeply conserved protein-protein interactions likely to be critical to ciliary function in humans. Examining human biological systems in the context of their level of conservation across species is a productive route to distinguish biological signal from noise. My preliminary data on three ciliated species confirm that this approach is capable of identifying conserved ciliary protein complexes. If a pair of proteins are found in physical contact in diverse species, having survived speciation events and gene loss, it can be predicted that this physical interaction is important. First, using mass spectrometry, we will experimentally detect protein complexes within the cilia across a spectrum of eukaryotic organisms, broadly defining conserved ciliary proteins. Second, we will integrate our own experimental data, some of which we have already collected, with outside data to construct a system-wide map of conserved ciliary protein complexes. These conserved ciliary protein complexes are strongly predicted to be functionally important in humans, and potentially involved in human birth defects. Finally, we will test a targeted subset of proteins predicted to associate with known birth defect proteins in vivo using Xenopus laevis embryos as a model system. We have four candidate genes in hand, and will consider candidates from the prior aims as appropriate. This project will lead to a map of critical conserved ciliary protein complexes, a future primary resource for research into diverse ciliopathies.

总结 纤毛病是一种不同类型的发育疾病，可以表现为肾脏，骨骼， 眼睛、心脏、生殖或精神功能。许多纤毛病变，包括Bardet-Biedl、Joubert和Meckel Gruber Syndrome是由睫状体的主要蛋白复合物缺陷引起的， 车厢至少有1,000种蛋白质与纤毛的生物发生和功能有关; 因此，纤毛蛋白组织的原理需要大规模的系统研究。我们目前 人类蛋白质复合物图谱虽然广泛，但只有中等范围的纤毛蛋白， 收集有针对性的实验数据。我将使用比较蛋白质组学分析的纤毛从多个 确定高度保守的蛋白质-蛋白质相互作用可能对纤毛功能至关重要， 人类在物种保护水平的背景下检查人类生物系统是一个 一种有效的区分生物信号和噪声的方法。我关于三种纤毛虫物种的初步数据 证实这种方法能够鉴定保守的纤毛蛋白复合物。如果一对蛋白质 在不同物种的身体接触中发现，在物种形成事件和基因丢失中幸存下来， 预测这种物理相互作用是重要的。首先，使用质谱，我们将实验性地 在真核生物的范围内检测纤毛内的蛋白质复合物， 保守纤毛蛋白。第二，我们将整合我们自己的实验数据，其中一些我们已经 已经收集，与外部数据构建保守的纤毛蛋白复合物的系统范围的地图。 这些保守的纤毛蛋白复合物被强烈预测在人类中具有重要的功能， 可能与人类出生缺陷有关最后，我们将测试一个有针对性的蛋白质子集， 使用非洲爪蟾胚胎作为模型系统，在体内与已知的出生缺陷蛋白相关联。我们有 四个候选基因在手，并将考虑候选人从以前的目标适当。该项目将 导致关键的保守纤毛蛋白复合物的地图，未来的主要资源，研究 各种纤毛病变