CORE--PROTEIN PURIFICATION FACILITY

蛋白质纯化中心将通过提供分离和纯化天然蛋白和重组蛋白的设施和技能，为所有项目提供支持。这些制备的原材料将由个别项目提供，并提供建议的分离方法。蛋白分离将由高级核心技术人员（50%的工作量）进行，并由核心领导（5%的工作量）监督。要求使用Pharmacia AKTA探索性蛋白质纯化系统，以保证时间和材料的有效利用。纯化后的蛋白质将通过分光光度法、sds凝胶电泳和特殊情况下的EPR光谱进行表征。核心将服务于所有项目，项目1（转铁蛋白的细胞铁摄取机制）、项目2（转铁蛋白受体的功能域）和项目5（转铁蛋白的结构、功能和动力学）使用最多，项目3 （SFT对转铁蛋白和非转铁蛋白结合铁的摄取）和项目4（铜在铁代谢中的作用）使用较少。

The Protein Purification Core will provide support to all projects by providing facilities and skills for isolating and purifying native and recombinant proteins. Raw materials for such preparations will be provided by the individual projects, along with suggested methods for the isolations. Protein isolations will be carried out the senior Core technician (50% effort), with supervision by Core leader (5% effort). A Pharmacia AKTA explorer Protein Purification System is requested to assume efficient use of time and materials. Purified proteins will by characterized by spectrophotometry, SDS-gel electrophoresis and, in special cases, by EPR spectroscopy. The Core will serve all projects, with heaviest use by Project 1 (Mechanisms of Cellular Iron Uptake From Transferrin, Project 2 (Functional Domains of the Transferrin Receptor), and Project 5 (Structure, Function and Dynamics in Transferrin), and less demanding use by Project 3 (Uptake of Transferrin and Non- transferrin Bound Fe by SFT) and Project 4 (Role of Copper in Iron Metabolism).