Crystallization and structural analysis of three transcriptional regulators from eubacteria and archaea: Aes. Mlc and TrmB

来自真细菌和古细菌的三种转录调节因子的结晶和结构分析：Aes。

• 批准号: 5409051
• 负责人: Professor Dr. Kay Diederichs
• 金额: --
• 依托单位: Fachbereich Biologie
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2003
• 资助国家: 德国
• 起止时间: 2002-12-31 至 2007-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/5409051?language=en
• 关键词: Crystallization; structural; analysis; three; transcriptional

We propose to crystallize and determine the three-dimensional structure of procaryotic transcription factors and proteins that are interacting with these regulators, that control their activity and couple this control to metabolic signal transduction chains. The frist protein is Aes from E. coli. It is an enzyme with acetyl esterase activity that exhibits homology to the family of hormone-sensitive lipases. The interest in this enzyme is based on its demonstrated interaction with MalT, the contral regulator of the E. coli maltose system. The second transcription regulator is Mlc, a global transcriptional repressor controlling the expression of the malT gene and in addition a number of genes encoding some specific proteins (for glucose and mannose) and all general enzymes of the E. coli phosphotransferase system (PTS). The interest in Mlc comes from its unusual mode of inactivation by sequestration to the PtsG transproter. We plan to determine the structure of Mlc als well as of the complex of Mlc with the soluble EIIB domain of PtsG. The third transcription regulator is TrmB, a sugar-specific transcription repressor controlling the genes encoding a high affinity and binding protein-dependent ABC transproter for trehalose and maltose in the hyperthermophilic archaeon Thermococcus litoralis. The interest in this regulator comes from the fact that is ist the first identified sugar specific regulator of archaea acting on a transcription machinery of eucaryotic relateness. The goal of this proposal is aimed at the understanding of a novel type of transcriptional regulation in which the regulator is controlled by direct protein-protein interaction.

我们建议结晶和确定原核生物转录因子和蛋白质的三维结构，这些转录因子和蛋白质与这些调控因子相互作用，控制它们的活性，并将这种控制耦合到代谢信号转导链上。第一个蛋白质是来自大肠杆菌的AES。它是一种具有乙酰酯酶活性的酶，表现出与激素敏感脂肪酶家族的同源性。对这种酶的兴趣是基于它与麦芽的相互作用，麦芽是大肠杆菌麦芽糖系统的控制调节器。第二个转录调节因子是MLC，它是一种全球转录抑制因子，控制MALT基因和一些编码某些特定蛋白(葡萄糖和甘露糖)的基因以及大肠杆菌磷酸转移酶系统(PTS)的所有通用酶的表达。对MLC的兴趣来自于它通过隔离到PtsG转运体的不寻常的失活模式。我们计划确定MLC ALS的结构以及MLC与PtsG的可溶性EIIB结构域的络合物的结构。第三个转录调节因子是TRMB，它是一种糖特异的转录抑制因子，控制着高温古生菌中海藻糖和麦芽糖的高亲和力和结合蛋白依赖的ABC转录子的编码基因。对这个调控因子的兴趣来自于这样一个事实，即它是第一个被发现的作用于真核相关转录机制的古生菌的糖特异性调控因子。这项建议的目的是为了理解一种新型的转录调控，其中调控因子由蛋白质-蛋白质直接相互作用控制。