STRUCTURE, FUNCTION AND DYNAMICS IN TRANSFERRIN

该项目的总体目标是发展一个详细的分子水平的理解金属结合转铁蛋白（TF）蛋白。这一目标将主要通过开发和应用基于激光的光谱工具与技术的组合来实现，以捕获或分离蛋白质的平衡和非平衡形式。站点和残基特定的构象探测Tf将主要使用紫外和可见光共振拉曼光谱和一个相对较新的镧系元素（III）为基础的电子振动发光光谱进行探索。这些振动光谱将补充吸收和荧光研究。该项目的这一部分的目标，包括确定功能不同的构象的Tf和功能重要的本地自由度的光谱签名。目的是能够探测局部相互作用作为全局构象变化的函数。光谱和动力学特性的Tf的特定构象将完成与添加一个新的多孔溶胶-凝胶为基础的封装方法的捕获和锁定的特定蛋白质结构。重点将放在探测载脂蛋白和金属结合形式的开放和封闭的确认Tf作为pH值的函数，阴离子，金属配体（螯合剂），转移受体，位点特异性突变和金属阳离子。

The overall objective of this project is to develop a detailed molecular level understanding of metal binding in transferrin (TF) proteins. This objective will be pursued primarily by developing and applying a combination of laser based spectroscopic tools in conjunction with techniques to trap or isolated both equilibrium and non-equilibrium forms of the protein. Site and residue specific conformational probing of Tf will be explored using primarily ultra violet and visible resonance Raman spectroscopy and a relatively new lanthanide (III) based vibronic luminescence spectroscopy. These vibrational spectroscopies will be supplemented with absorption and fluorescence studies. Objectives of this part of the project include identifying spectroscopic signatures both for functionally distinct conformations of Tf and for functionally important local degrees of freedom. The intent is to be able to probe the local interactions as a function of changes in the global conformations. Spectroscopic and kinetic characterization of specific conformations of Tf will be accomplished with the addition of a new porous sol-gel based encapsulation method of trapping and locking in of specific protein structures. An emphasis will be placed on probing the apo and metal bound forms of both the open and closed confirmations of Tf as a function of pH, anions, metal ligands (chelators), transferring receptor, site-specific mutations and metal cations.