CORE--TECHNICAL CORE

核心--技术核心

基本信息

批准号：
7537249
负责人：
WOLFGANG H FISCHER
金额：
$ 55.07万
依托单位：
SALK INSTITUTE FOR BIOLOGICAL STUDIES
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-12-01 至 2008-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7537249
关键词：
ACVR1 gene Activins Affinity Amino Acids Animals Antibodies Antibody Formation Antigens Binding Binding Proteins Biological Blocking Antibodies Blood capillaries C-terminal Carrier Proteins Cells Chemicals Chimeric Proteins Chinese Hamster Ovary Cell Circular Dichroism Complementary DNA Complex Consult Controlled Study Coupling Development Electrophoresis Endoglin Equipment Estradiol Event Extracellular Domain Family member Follicle Stimulating Hormone Follistatin Gel Gene Proteins Generations Glutathione S-Transferase Goals Gonadal Steroid Hormones Growth and Development function High Pressure Liquid Chromatography Human Resources Immune Sera Immunoassay Immunoblotting Immunohistochemistry Immunoprecipitation In Vitro Individual Lead Ligands Liquid substance Mass Spectrum Analysis Measurement Membrane Methods Molecular Conformation Mus NMR Spectroscopy Nature Numbers Pattern Peptide Synthesis Peptides Phase Physical condensation Physiological Pituitary Gland Pituitary Hormones Plasma Post-Translational Protein Processing Preparation Principal Investigator Procedures Production Progesterone Proteins Proteoglycan Protocols documentation Quality Control Radioimmunoassay Reagent Recombinant Activin Recombinant Proteins Recombinants Regulation Research Personnel Role Sampling Sepharose Sequence Analysis Services Signal Transduction Solid Standardization Steroids Structure Structure-Activity Relationship System TGF beta type III receptor Techniques Testosterone Training UMOD gene Update activin A activin B capillary cell growth cost cross reactivity design disulfide bond immunoaffinity chromatography improved inhibin inhibin B inhibin receptor instrumentation interest method development novel polypeptide programs protein expression receptor receptor binding reproductive function research study sex synthetic peptide tissue culture urinary gonadotropin fragment

项目摘要

This Program Project focuses on the role of activins and inhibins and their membrane-bound receptors, soluble binding proteins and modulators in the regulation of reproductive function, cellular growth and development. The primary focus of this Program is to characterize the physical interactions and molecular events that lead to signal propagation by activin and inhibin. The second focus is to explore the physiologic and pathophysiologic significance of these molecules at the cellular and system levels and to study the control of protein expression and secretion as well as modes of action. The Technical Core (Core Component B) provides vital services, including protein expression, peptide synthesis, peptide/protein characterization, and generation of antibodies, needed by the Projects of this Program to carry out structural, functional and mechanistic studies. Large quantities of recombinant activin, inhibin and betaglycan fragments, essential for the structural and biological studies, will be expressed by the Core. Peptides for generation of antisera and for specific blockers of antibodies will be synthesized and purified by the Core. Synthetic peptides and recombinant proteins generated by the Core, additional recombinant proteins produced by individual Projects, and novel proteins isolated by this Program will be characterized. Posttranslational modifications of ligands and/or receptors will be identified. The analysis of polypeptides will be achieved using a number of techniques including HPLC, mass spectroscopy, NMR and automated Edman degradation. Synthetic peptides conjugated to carrier proteins will be used as immunogens to develop antibodies directed towards gene products predicted from cDNA sequences. Alternatively, proteins produced using in vitro cell expression systems will be injected directly into host animals for production of antisera. To reduce cross-reactivity with antigens other than the protein of interest, selected antisera will be immunoaffinity purified when necessary. Core B will additionally provide immunoassays of pituitary gonadotropic hormones and sex steroids for physiologic and pharmalogic studies proposed. The reagents and services provided require specialized equipment and highly trained personnel which can be most efficiently consolidated in a Core. In addition, the integrated nature of the Core assures cost efficiency, standardization of protocols and facilitates quality control.

该程序项目的重点是激活素和抑制素及其膜结合的受体，可溶性结合蛋白和调节剂在生殖功能，细胞生长和发育中的作用。该程序的主要重点是表征物理相互作用和分子事件，从而导致激活素和抑制素的信号传播。第二个重点是探索这些分子在细胞和系统水平上的生理和病理生理意义，并研究蛋白质表达和分泌以及作用方式的控制。技术核心（核心成分B）提供了重要服务，包括蛋白质表达，肽合成，肽/蛋白质表征和抗体的产生，该计划的项目需要进行结构，功能和机械研究。大量的重组激活素，抑制素和βlycan片段对于结构和生物学研究至关重要，将由核心表达。用于生成抗血清和特定抗体阻滞剂的肽将通过核心合成和纯化。由核心产生的合成肽和重组蛋白，由单个项目产生的其他重组蛋白以及该程序分离的新型蛋白质。将确定配体和/或受体的翻译后修饰。使用多种技术（包括HPLC，质谱，NMR和自动化的Edman降解）来实现多肽的分析。与载体蛋白结合的合成肽将用作免疫原，以开发针对cDNA序列预测的基因产物的抗体。或者，使用体外细胞表达系统产生的蛋白质将直接注入宿主动物以生产抗血清。为了降低与感兴趣的蛋白质以外的抗原的交叉反应性，必要时会纯化选定的抗血清。核心B将提供垂体性促性腺激素激素和性类固醇的免疫测定，用于生理和药物研究。提供的试剂和服务需要专门的设备和受过良好训练的人员，这些设备可以最有效地巩固核心。此外，核心的综合性质可确保成本效率，协议的标准化并促进质量控制。