Mapping the blood cell protein complexosome

绘制血细胞蛋白复合体图谱

基本信息

批准号：
9160716
负责人：
Andrew EMILI
金额：
$ 25.05万
依托单位：
UNIVERSITY OF TEXAS AT AUSTIN
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-07-01 至 2019-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9160716
关键词：
Affect Anemia Aplastic Anemia Binding Proteins Biochemical Biological Assay Biology Blood Blood Cells Cell Line Cell Nucleus Cell Shape Cell physiology Cells Complex Computer Simulation Cytoplasmic Protein Data Set Defect Development Disease Erythroblasts Erythrocytes Excision Foundations Fractionation Future Grant Hematological Disease Hematopoietic Hemoglobin Human Immunoprecipitation Information Networks Kidney Failure Label Lead Longevity Macromolecular Complexes Mammals Maps Mass Spectrum Analysis Measures Megaloblastic Anemia Membrane Membrane Proteins Metabolic Methods Modeling Molecular Multipotent Stem Cells Multiprotein Complexes Mus Organelles Oryctolagus cuniculus Pathway interactions Pilot Projects Play Production Property Protein Biochemistry Proteins Proteomics Recombinants Recovery Role Shapes Shotguns System Techniques Testing Transfection Work base cell type comparative gene function immortalized cell insight macromolecule network models new technology novel oxygen transport progenitor protein complex protein expression protein protein interaction research study response transcriptome sequencing wasting

项目摘要

ABSTRACT Erythrocytes (red blood cells; RBCs) and their progenitors express distinct proteins, which underlie their unique biology, and which provide a molecular basis for many blood diseases, including diverse anemias, such as those arising during renal failures as a result of low red blood cell production and lifespan. Importantly, mammalian RBCs lack nuclei and other major organelles, and hence neither transcriptional profiling by RNA- sequencing nor recombinant transfection—powerful techniques in other cell types—can be used to reveal RBC gene functions and pathways. Proteomics methods, in contrast, allow for a detailed analysis of RBC proteins, and pioneering studies have revealed that RBCs, while dominated by hemoglobin (98%), express on the order of 1,500 to 2,000 distinct proteins. Many of these proteins play critical roles in erythrocyte function, including key metabolic and bioenergy roles, and cytoskeletal roles in controlling RBC cell shape. More than 500 proteins in RBCs are of entirely unknown function. A fundamental question in RBC biology is thus how all of these proteins work together to support proper RBC function and development. Building deep mechanistic understanding of RBC biology requires accurately delineating the precise membership of protein complexes specific to RBCs, as these carry out key functions unique to these cells. We propose to perform the first systematic, global exploration of native protein-protein interactions (PPIs) in RBCs, using a powerful new technology to examine those interactions directly among endogenous proteins in human and other mammalian RBCs. Our proposed experiments combine protein biochemistry, quantitative mass spectrometry proteomics and integrative computer modeling to reliably define the extended PPI networks and multiprotein complexes native to RBCs, helping to lay rich new mechanistic foundations for interpreting RBC biology. By the end of this grant, we will have performed nearly 2,000 mass spectrometry experiments on native protein complexes isolated from primary RBCs and their progenitors, defining the RBC interactome, including both shared and novel protein complexes, to an unprecedented degree. As a result of this work, RBCs will be the first primary human cell type with a near complete map of stable protein complexes, giving new insights into erythrocyte biology and development, and laying the foundation for future attempts to intervene, chemically or genetically, in diseases affecting these critical cells.

抽象的红细胞（红细胞； RBC）及其祖细胞表达不同的蛋白质，这是其独特的基础生物学，这为许多血液疾病提供了分子基础由于红细胞的产生和寿命低，在肾脏失败期间出现的人。哺乳动物RBC缺乏核和其他主要细胞器，而Hense均未通过RNA-进行转录分析测序或重组转染 - 其他细胞类型中的强大技术 - 可用于RBEL RBC 相反，基因功能和途径。开创性的研究使加拿大皇家银行（RBC）陶醉，而由血红蛋白（98％）主导 1,500至2,000个不同的蛋白质。关键的代谢和生物能源作用，以及控制RBC细胞形状的细胞骨骼作用 RBC中的蛋白质具有输入未知功能。这些蛋白质共同支持适当的RBC功能和开发了解RBC生物学要求准确地描述了蛋白质复合物的精确成员特定于RBC，因为执行密钥功能是分区的独特功能。使用强大的新的新的RBC中，系统的全球探索天然蛋白质蛋白质相互作用（PPI）（PPI）在人和其他哺乳动物中直接检查内源蛋白之间的这些相互作用的技术 RBC。我们的支撑实验结合了蛋白质生物化学以及可靠的集成计算机建模定义扩展的PPI网络和多蛋白复合物原产于RBC，在此结束时，有助于解释RBC生物学的新机构基础。 Grant，我们将对天然蛋白质复合物进行近2,000次大型测量实验从主要的RBC和祖细胞中分离出来，定义RBC Interactome，包括共享新型蛋白质复合物，由于这项工作而成为前所未有的degree。人类细胞类型，近乎完整的稳定蛋白质复合物图，为红细胞提供了新的见解生物学和发展，并为未来的化学或遗传干预尝试奠定基础在疾病中，对这些批判性细胞感兴趣。