Illumination of the Structure and Function of PptA, a Novel Type of Polyphosphate Granule Attached Protein in Organelle-like Polyphosphatosomes of Ralstonia eutropha H16

真养罗尔斯通氏菌 H16 细胞器样多磷酸体中新型多磷酸盐颗粒附着蛋白 PptA 的结构和功能的阐明

• 批准号: 400441076
• 负责人: Professor Dr. Dieter Jendrossek
• 金额: --
• 依托单位: Institut für Mikrobiologie (IMB)
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2018
• 资助国家: 德国
• 起止时间: 2017-12-31 至 2020-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/400441076?language=en
• 关键词: Illumination; Structure; Function; PptA; Novel

In this project we will determine the structure and function of PptA, a protein of R. eutropha that is characterized by an ubiquitously occurring (in all kingdoms of life) CHAD (conserved histidine alpha-helical) domain at the edge of P- and N-metabolism. PptA has two remarkable proterties: (i) it´s CHAD domain confers a specific binding to polyP granules in a species independent manor and (ii) it interacts with the key enzyme of N-metabolism, glutamine synthetase GlnA1. Here, we will determine whether arginines of conserved sequence motifs (isoelectric point of PptA is 11.5!) or other residues of PptA, that are conserved in CHAD proteins, are responsible for interaction with the polyanion polyP and/or GlnA1 (IEP 5.4). The strength and stoichiometry of the PptA-GlnA1, PptA-polyP and GlnA1-polyP interaction will be determined by isothermic colometric titration (ITC) and/or surface plasmon resonance (SPR). The influence of the PptA-GlnA1 interaction on the oligomeric state and activity and of GlnA1 in the presence of different effector molecules will be determined by light scattering technique.

本项目将研究红曲霉PptA蛋白的结构和功能。真养型，其特征在于在P-和N-代谢的边缘处普遍存在（在所有生命界中）CHAD（保守的组氨酸α-螺旋）结构域。PptA具有两个显著的特性：（i）其CHAD结构域以物种独立的方式赋予聚P颗粒特异性结合;（ii）其与N-代谢的关键酶谷氨酰胺合成酶GlnA 1相互作用。在这里，我们将确定是否保守序列基序（PptA的等电点为11.5！）或在CHAD蛋白中保守的PptA的其他残基负责与聚阴离子polyP和/或GlnA 1相互作用（IEP 5.4）。PptA-GlnA 1、PptA-polyP和GlnA 1-polyP相互作用的强度和化学计量将通过等温比色滴定（ITC）和/或表面等离子体共振（SPR）来确定。将通过光散射技术测定PptA-GlnA 1相互作用对寡聚体状态和活性的影响以及在不同效应分子存在下GlnA 1的影响。