Kondakova Tatiana, Merlet-Machour Nadine, Chapelle Manuel, Preterre David, Dionnet Frédéric, Feuilloley Marc, Orange Nicole, Duclairoir Poc Cécile
Laboratory of Microbiology Signals and Microenvironment (LMSM) EA4312, Normandy Univ., Univ. Rouen, 55 rue St Germain, 27000 Evreux, France; Aerothermic and Internal Combustion Engine Technological Research Center (CERTAM), 1 Rue Joseph Fourier, 76800 Saint Etienne du Rouvray, France.
Team Modified to Surface and Interface Analysis (SIMA), UMR 6014 COBRA, Normandy Univ., Univ. Rouen, 55 rue St Germain, 27000 Evreux, France.
Res Microbiol. 2015 Jan;166(1):1-8. doi: 10.1016/j.resmic.2014.11.003. Epub 2014 Dec 2.
Lipids are major functional components of bacterial cells that play fundamental roles in bacterial metabolism and the barrier function between cells and the environment. In an effort to investigate the bacterial lipidome, we adopted a protocol using MALDI-TOF MS imaging coupled to HPTLC to screen a large number of phospholipid classes in a short span of time. With this method, phospholipids of airborne Pseudomonas fluorescens MFAF76a were visualized and identified in sample extracts (measurement accuracy below 0.1 Da, phospholipid identification by means of four characteristic fragment peaks). Via this technique, the P. fluorescens lipidome was shown to comprise three major lipid classes: phosphatidylethanolamine, phosphatidylglycerol and phosphatidylcholine. The protocol described herein is simple, rapid and effective for screening of bacterial phospholipid classes. The remarkable presence of a eukaryotic phospholipid, phosphatidylcholine, was observed in P. fluorescens MFAF76a. This lipid is known to play a role in bacteria-host interactions and had not been known to be found in P. fluorescens cells.
脂质是细菌细胞的主要功能成分,在细菌代谢以及细胞与环境之间的屏障功能中发挥着重要作用。为了研究细菌脂质组,我们采用了一种将基质辅助激光解吸电离飞行时间质谱成像(MALDI-TOF MS成像)与高效薄层层析(HPTLC)相结合的方法,以便在短时间内筛选大量磷脂类别。通过这种方法,在样品提取物中可视化并鉴定了空气传播的荧光假单胞菌MFAF76a的磷脂(测量精度低于0.1 Da,通过四个特征性碎片峰鉴定磷脂)。通过该技术,荧光假单胞菌脂质组显示包含三类主要脂质:磷脂酰乙醇胺、磷脂酰甘油和磷脂酰胆碱。本文所述方法对于筛选细菌磷脂类别而言简单、快速且有效。在荧光假单胞菌MFAF76a中观察到了真核磷脂磷脂酰胆碱的显著存在。已知这种脂质在细菌与宿主的相互作用中发挥作用,并且此前从未在荧光假单胞菌细胞中发现过。