CORE--PROTEIN PURIFICATION FACILITY

蛋白质纯化核心将通过提供分离和纯化天然和重组蛋白质的设施和技能，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，以及建议的隔离方法。蛋白质分离将由资深核心技术员(50%的努力)在核心领导的监督下(5%的努力)进行。需要一套Pharmacia Akta探索者蛋白质纯化系统，以实现时间和材料的高效利用。纯化的蛋白质将通过分光光度法、十二烷基硫酸钠-凝胶电泳法和特殊情况下的电子顺磁共振波谱进行表征。核心将服务于所有项目，使用最多的是项目1(转铁蛋白的细胞铁吸收机制)、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)，需求较少的项目3(通过SFT吸收转铁蛋白和非转铁蛋白结合的铁)和项目4(铜在铁代谢中的作用)。

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).