CORE--TECHNICAL CORE

本项目主要研究激活素和抑制素及其膜结合受体、可溶性结合蛋白和调节剂在调节生殖功能、细胞生长和发育中的作用。该计划的主要重点是表征激活素和抑制素导致信号传播的物理相互作用和分子事件。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，并研究其对蛋白质表达和分泌的控制及其作用方式。技术核心（核心组件B）提供关键服务，包括蛋白表达、肽合成、肽/蛋白表征和抗体生成，这些都是本计划项目开展结构、功能和机制研究所需的。大量的重组激活素、抑制素和β多糖片段，对结构和生物学研究至关重要，将由Core表达。用于生成抗血清和特异性抗体阻滞剂的肽将由Core合成和纯化。将对Core生成的合成肽和重组蛋白、个别项目生成的其他重组蛋白以及本项目分离的新蛋白进行表征。将鉴定配体和/或受体的翻译后修饰。多肽的分析将使用包括HPLC、质谱、NMR和自动Edman降解在内的许多技术来实现。与载体蛋白结合的合成肽将作为免疫原用于开发针对cDNA序列预测的基因产物的抗体。另外,

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.