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：
8555145
负责人：
Frank Leo Conlon
金额：
$ 24.62万
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/8555145
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）：摘要该提案的目标是开发和优化用于非洲爪蟾定向和定量蛋白质组学方法的技术和试剂。具体来说，我们提出了一系列基于三种蛋白质的可行性研究，即T-box转录因子TBX5、锌指转录因子CASZ1和非受体蛋白磷酸酶SHP2。我们关注这些蛋白质是基于它们在序列、表达和功能方面的进化保守性、它们的细胞定位，以及这些蛋白质是致病性的或与人类疾病状态相关的观察结果：霍尔特-奥拉姆综合征 (HOS) 患者、一种也影响心脏和四肢的疾病，与 Tbx5 的错义突变有关，全基因组关联研究表明 Casz1 在高血压和努南综合征（一种也影响心脏和四肢的疾病）患者中发挥作用。肢发育通常与 Shp-2 的错义突变有关。尽管 TBX5、CASZ1 和 SHP-2 在发育和人类疾病中发挥着至关重要的作用，但这些蛋白质在体内发挥作用的分子机制仍有待确定。为了解决这些问题并建立一套在爪蟾中使用定向蛋白质组学方法的通用技术，我们建议 1) 在爪蟾中建立二元转基因系统，用于分离蛋白质和蛋白质-DNA 复合物，2) 建立技术，用于爪蟾定向和定量蛋白质组学方法的试剂和方案。