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

这个项目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的E.ColiEntf。该蛋白由C、A、T和TE四个结构域组成 作为链延长和终止模块，制造和释放肠结合蛋白铁载体。我们 感兴趣的是结构域如何协作合成Enterobactin，以及HS-Pantetheinyl-P如何 与TE结构域结合的基团影响结构域的相互作用，以及这可能如何与组装- 蛋白质的线功能。多结构域蛋白质是大分子核磁共振研究的模型系统 蛋白质类 具体目标是： 具体目标1..142 kDa的36 kDa A-T双域片段的完整结构测定 Entf 特定目标2以T为中心的In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其对接 目的3.Entf A-T 70 kDa双域结构的研究 4.明确目标。HS-Pantetheinyl-P修复基(同位素标记或自旋标记)的相互作用 )在顺式和反式(C，A，TE)中具有不同结构域的EntB和Entf中的两个T结构域 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