STRUCTURE, FUNCTION AND DYNAMICS IN TRANSFERRIN

该项目的总体目标是在分子水平上对转铁蛋白（TF）中的金属结合进行详细的了解。这一目标将主要通过开发和应用基于激光的光谱工具结合技术来捕获或分离蛋白质的平衡和非平衡形式来实现。将主要使用紫外和可见共振拉曼光谱以及相对较新的基于镧系元素（III）的振动发光光谱来探索Tf的位点和残基特定构象探测。这些振动光谱将补充吸收和荧光研究。这部分项目的目标包括识别Tf功能上不同构象和功能上重要的局部自由度的光谱特征。其目的是能够探测局部相互作用作为全局构象变化的函数。通过添加一种新的多孔溶胶-凝胶包封方法来捕获和锁定特定的蛋白质结构，将完成Tf特定构象的光谱和动力学表征。重点将放在探测载脂蛋白和金属结合形式的开放和关闭确认的Tf作为一个函数的pH，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.