Direct and Quantitative Proteomic Approaches in Xenopus

非洲爪蟾的直接定量蛋白质组学方法

基本信息

批准号：
8710298
负责人：
Frank Leo Conlon
金额：
$ 18.97万
依托单位：
UNIV OF NORTH CAROLINA CHAPEL HILL
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-08-01 至 2015-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8710298
关键词：
Acute Lymphocytic Leukemia Acute Myelocytic Leukemia Address Adult Affect Affinity Amino Acid Sequence Animals Antibodies Biological Models Biotin Boxing Buffers Castor Collaborations Communities Complex Core Facility Cost Savings DNA Databases Deposition Development Disease Embryo Enzymes Feasibility Studies Future Generations Genes Genome Goals Heart Holt Oram syndrome Hypertension Immunoprecipitation Juvenile Myelomonocytic Leukemia Label Limb Development Limb structure Link Liquid Chromatography Mass Spectrum Analysis Messenger RNA Methodology Molecular Mutate Mutation Noonan Syndrome PTPN11 gene Patients Peptide Sequence Determination Peptides Post-Translational Protein Processing Procedures Protein Databases Protein phosphatase Proteins Proteomics Protocols documentation Reagent Resources Role Series Specificity Staging Streptavidin Syndrome System Tadpoles Technology Testing Time Tissues Transgenic Animals Transgenic Organisms Xenopus Xenopus Proteins Xenopus laevis Zinc Fingers base cardiogenesis chromatin immunoprecipitation congenital heart disorder cost egg embryo tissue genome wide association study human disease in vivo novel protein complex protein purification public health relevance receptor research study sperm cell tandem mass spectrometry transcription factor

项目摘要

DESCRIPTION (provided by applicant): ABSTRACT The goal of this proposal is to develop and optimize the technologies and reagents for use of directed and quantitative proteomic approaches in Xenopus. Specifically, we propose a series of feasibility studies based on three proteins, the T-box transcription factors TBX5, the zinc finger transcription factor CASZ1, and the non-receptor protein phosphatase SHP2. We have focused on these proteins based on their evolutionary conservation in sequence, expression, and function, their cellular localization, and the observation that these proteins are causative, or have association with human disease states: patients with Holt-Oram syndrome (HOS), a disease which also affects the heart and limbs, is associated with mis-sense mutations in Tbx5, genome wide association studies have implied a role for Casz1 in hypertension and high blood pressure, and patients with Noonan syndrome, a disease that also affects heart and limb development, frequently is associated with mis-sense mutations in Shp-2. Despite the crucial role for TBX5, CASZ1 and SHP-2 in development and human disease, the molecular mechanisms by which these proteins act in vivo remains to be established. To address these issues and to establish a set of general technologies for the use of directed proteomics approaches in Xenopus we propose to 1) establish a binary transgenic system in Xenopus for the isolation of protein and protein-DNA complexes and 2) establish the technologies, reagents and protocols for directed and quantitative proteomic approaches in Xenopus.

描述（由申请人提供）：摘要该提案的目的是开发和优化Xenopus中使用定向和定量蛋白质组学方法的技术和试剂。具体而言，我们提出了一系列基于三种蛋白质，T-box转录因子TBX5，锌指转录因子Casz1和非受体蛋白磷酸酶SHP2的可行性研究。我们基于这些蛋白质的进化保守性，表达和功能，其细胞定位以及这些蛋白质是病因或与人类疾病有关联的观察到：Holt-Oram综合征（HOS），，HOLT-ORAM综合征（HOS），一种也影响心脏和四肢的疾病与TBX5中的错误敏感突变有关，基因组广泛的关联研究暗示了Casz1在高血压和高血压中的作用，以及Noonan综合征的患者，这种疾病也会影响心脏和心脏和血压肢体发育经常与SHP-2中的错误敏感突变有关。尽管TBX5，CASZ1和SHP-2在发育和人类疾病中起着至关重要的作用，但这些蛋白质在体内作用的分子机制仍有待确定。为了解决这些问题并建立一组通用技术，用于使用爪诺邦中的定向蛋白质组学方法，我们建议1）在Xenopus中建立一个二元转基因系统，用于隔离蛋白质和蛋白质-DNA复合物，以及2）建立该技术，2）爪蟾中定向和定量蛋白质组学方法的试剂和方案。