A phosphatidic acid-binding protein is important for lipid homeostasis and adaptation to anaerobic biofilm conditions in Pseudomonas aeruginosa.
A phosphatidic acid-binding protein is important for lipid homeostasis and adaptation to anaerobic biofilm conditions in Pseudomonas aeruginosa.
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DOI：
10.1042/bcj20180257
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发表时间：
2018-06
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通讯作者：
Maike K Groenewold;Marco Massmig;Stefanie Hebecker;Linna Danne;Zofia Magnowska;M. Nimtz;F. Narberhaus;D. Jahn;D. Heinz;L. Jänsch;J. Moser
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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
定量铜绿假单胞菌蛋白质组学方法揭示了在囊性纤维化肺条件下生长的厌氧生物膜中迄今未表征的蛋白质PA 3911的丰度增加。ORF PA 3911的生理相关性阿利亚使用表型微阵列实验来证明。突变株在抗菌剂（米诺环素，萘夫西林，苯唑西林，氯霉素和甲砜霉素）的存在下表现出增加的敏感性，增强抽搐运动性和显着受损的生物膜形成。PA 3911是铜绿假单胞菌中的可溶性细胞质蛋白。在蛋白质-脂质重叠实验中，纯化的PA 3911特异性结合磷脂酸（PA），磷脂代谢的中心枢纽。结构指导的定点突变被用来探索PA 3911的配体结合腔。Leu 56、Leu 58、Val 69和Leu 114的蛋白质变体显示出损害PA相互作用。比较鸟枪脂质组学方法证明了铜绿假单胞菌在脂质头基团和脂肪酸水平上对厌氧条件的多方面反应。在厌氧和/或需氧条件下，PA 3911突变株中的脂质稳态相对于溶血磷脂酰胆碱、磷脂酰胆碱和甘油二酯不平衡。新发现的PA结合蛋白对脂质稳态和相关的宏观表型铜绿假单胞菌的影响进行了讨论。
A quantitative Pseudomonas aeruginosa proteomics approach revealed increased abundance of the so-far uncharacterized protein PA3911 in anaerobic biofilms grown under conditions of the cystic fibrosis lung. Physiological relevance of ORF PA3911 was demonstrated, inter alia, using phenotype microarray experiments. The mutant strain showed increased susceptibility in the presence of antimicrobials (minocycline, nafcillin, oxacillin, chloramphenicol and thiamphenicol), enhanced twitching motility and significantly impaired biofilm formation. PA3911 is a soluble, cytoplasmic protein in P. aeruginosa In protein-lipid overlay experiments, purified PA3911 bound specifically to phosphatidic acid (PA), the central hub of phospholipid metabolism. Structure-guided site-directed mutagenesis was used to explore the proposed ligand-binding cavity of PA3911. Protein variants of Leu56, Leu58, Val69 and Leu114 were shown to impair PA interaction. A comparative shotgun lipidomics approach demonstrated a multifaceted response of P. aeruginosa to anaerobic conditions at the lipid head group and fatty acid level. Lipid homeostasis in the PA3911 mutant strain was imbalanced with respect to lysophosphatidylcholine, phosphatidylcholine and diacylglycerol under anaerobic and/or aerobic conditions. The impact of the newly identified PA-binding protein on lipid homeostasis and the related macroscopic phenotypes of P. aeruginosa are discussed.