NMR Studies of EntF-EntB Components of E.Coli Enterobactin Synthetase/Component 4

本研究的总体目标是了解非核糖体多肽中结构域之间的相互作用 合成酶模块，使用142 kDa E. coli EntF.该蛋白由C、A、T和TE四个结构域组成 其作为链延长和终止模块以制造和释放肠杆菌素铁载体。我们 感兴趣的是这些结构域如何合作合成肠杆菌素，以及HS-泛酰巯基乙胺-P 与TE结构域结合的基团影响结构域的相互作用，以及这可能与组装有关的方式。 蛋白质的线性功能。多结构域蛋白质是用于开发大规模NMR的模型系统。 蛋白 具体目标是： 具体目标1.. 142 kDa的36 kDa A-T双结构域片段的完全结构测定 EntF 具体目标2以T为中心的顺式C和A结构域的结构和相互作用的研究 结构域支架以及这些伙伴蛋白如何与之对接 第三章. EntF的A-T 70 kDa双结构域的结构研究 具体目标4。. HS-泛酰巯基乙胺基-P辅基（同位素标记或自旋标记的）的相互作用 ）在EntB和EntF中的两个T结构域上，在未酰化的 和酰化形式

The overall goal of this project is to learn about the interaction of domains in non-ribosomal polypeptide synthetase modules, using the 142 kDa E. coli EntF. This protein consists of four domains, C, A, T and TE that act as chain elongation and termination module to make and release the enterobactin siderophore. We are interested in how the domains cooperate to synthesize enterobactin, and how the HS-pantetheinyl-P group bound to the TE domain affects the interaction of domains, and how this may relate to the assembly- line function of the protein. The multidomain protein is a model system for development of NMR for large proteins The specific aims are: Specific Aim 1.. Complete structure determination of the 36 kDa A-T didomain fragment of the 142kDa EntF Specific Aim 2 Studies of structures and interactions of the in cis C and A domains centering on the T domain scaffold and how these partner proteins dock with it SpecificAim 3. Studies of the structure of the A-T 70 kDa didomain of EntF Specific Aim 4. . Interaction of the HS-pantetheinyl-P prosthetic group (isotopically labeled or spin-labeled )on the two T domains in EntB and EntF with different domains in cis and in trans (C, A, TE) in unacylated and acylated forms