tRNA-abhängige Biosynthesis of Phospholipids in Pseudomonas aeruginosa

铜绿假单胞菌中依赖 tRNA 的磷脂生物合成

• 批准号: 162323379
• 负责人: Professor Dr. Dirk Heinz
• 金额: --
• 依托单位: Helmholtz-Zentrum für Infektionsforschung (HZI)
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2009
• 资助国家: 德国
• 起止时间: 2008-12-31 至 2016-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/162323379?language=en
• 关键词: tRNA; abh; ngige; Biosynthesis; Phospholipids

Bacterial membranes undergo permanent changes in their composition in response to varying environmental conditions to efficiently fulfil their biological tasks. For this purpose lipids are covalently modified for instance by adding lysine onto phosphatidylglycerol (PG), thereby altering the net negative charge of the membrane. This lipid modification mediates resistance to cationic antimicrobial peptides (CAMPs) in Gram positive bacteria. Recently we discovered the tRNA-dependent addition of alanine to PG with the formation of a new phospholipid alanyl-phosphatidylglycerol (A-PG) in the Gram negative pathogen Pseudomonas aeruginosa mediating resistance against two positively charged antibacterial agents, a β-lactam antibiotic, high concentrations of H+ and lactate. Here we want to elucidate the tRNA-dependent enzyme mechanism and the mode of substrate recognition for the corresponding alanyl-phosphatidylglycerol synthase (A-PGS) using purified recombinant P. aeruginosa enzyme and advanced biochemical and biophysical techniques. A crystal structure will provide the understanding of the involved structure-function relationships. Novel biochemical principles of general importance are expected. Furthermore the physiological and biophysical role of A-PG formation versus L-PG formation will be elucidated. Finally the influence of alkaline conditions on the lipid composition will be analyzed analogously.

细菌膜在其组成中发生永久性变化，以响应不同的环境条件，以有效地完成其生物学任务。为此目的，例如通过将赖氨酸添加到磷脂酰甘油（PG）上来共价修饰脂质，从而改变膜的净负电荷。这种脂质修饰介导革兰氏阳性菌对阳离子抗菌肽（CAMP）的抗性。最近，我们发现在革兰氏阴性病原体铜绿假单胞菌中丙氨酸对PG的tRNA依赖性加成，形成新的磷脂丙氨酰磷脂酰甘油（A-PG），介导对两种带正电荷的抗菌剂（β-内酰胺抗生素、高浓度H+和乳酸盐）的抗性。本研究利用纯化的重组铜绿假单胞菌酶和先进的生物化学和生物物理技术，阐明了相应的丙氨酰磷脂酰甘油合成酶（A-PGS）的tRNA依赖性酶机制和底物识别模式。晶体结构将提供对所涉及的结构-功能关系的理解。新的生物化学原则的普遍重要性。此外，将阐明A-PG形成与L-PG形成的生理和生物物理作用。最后将类似地分析碱性条件对脂质组成的影响。

项目成果

Structures of two bacterial resistance factors mediating tRNA-dependent aminoacylation of phosphatidylglycerol with lysine or alanine

• DOI: 10.1073/pnas.1511167112
• 发表时间: 2015-08-25
• 期刊: PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
• 影响因子: 11.1
• 作者: Hebecker, Stefanie;Krausze, Joern;Moser, Juergen
• 通讯作者: Moser, Juergen

Alanyl‐phosphatidylglycerol synthase: mechanism of substrate recognition during tRNA‐dependent lipid modification in Pseudomonas aeruginosa

丙氨酰磷脂酰甘油合酶：铜绿假单胞菌 tRNA 依赖性脂质修饰过程中底物识别的机制

• DOI: 10.1111/j.1365-2958.2011.07621.x
• 发表时间: 2011
• 期刊: Molecular Microbiology
• 影响因子: 3.6
• 作者: Hebecker;Arendt;Heinemann;Tiefenau
• 通讯作者: Tiefenau

A phosphatidic acid-binding protein is important for lipid homeostasis and adaptation to anaerobic biofilm conditions in Pseudomonas aeruginosa.

• DOI: 10.1042/bcj20180257
• 发表时间: 2018-06
• 期刊: The Biochemical journal
• 作者: Maike K Groenewold;Marco Massmig;Stefanie Hebecker;Linna Danne;Zofia Magnowska;M. Nimtz;F. Narberhaus;D. Jahn;D. Heinz;L. Jänsch;J. Moser

Resistance Phenotypes Mediated by Aminoacyl-Phosphatidylglycerol Synthases

• DOI: 10.1128/jb.06576-11
• 发表时间: 2012-03-01
• 期刊: JOURNAL OF BACTERIOLOGY
• 影响因子: 3.2
• 作者: Arendt, Wiebke;Hebecker, Stefanie;Moser, Juergen
• 通讯作者: Moser, Juergen