Righi Valeria, Constantinou Caterina, Kesarwani Meenu, Rahme Laurence G, Tzika Aria A
Nuclear Magnetic Resonance Surgical Laboratory, Department of Surgery, Division of Burns, Massachusetts General Hospital and Shriners Burns Institute, Harvard Medical School, Boston, MA 02114, USA ; Athinoula A. Martinos Center for Biomedical Imaging, Department of Radiology, Massachusetts General Hospital, Harvard Medical School, Boston, MA 02114, USA ; Department for Life Quality, University of Bologna, Rimini 47921, Italy.
Nuclear Magnetic Resonance Surgical Laboratory, Department of Surgery, Division of Burns, Massachusetts General Hospital and Shriners Burns Institute, Harvard Medical School, Boston, MA 02114, USA ; Athinoula A. Martinos Center for Biomedical Imaging, Department of Radiology, Massachusetts General Hospital, Harvard Medical School, Boston, MA 02114, USA.
Biomed Rep. 2013 Sep;1(5):707-712. doi: 10.3892/br.2013.148. Epub 2013 Jul 22.
(PA) is a pathogenic gram-negative bacterium that is widespread in nature, inhabiting soil, water, plants and animals. PA is a prevalent cause of deleterious human infections, particularly in patients whose host defense mechanisms have been compromised. Metabolomics is an important tool used to study host-pathogen interactions and to identify novel therapeutic targets and corresponding compounds. The aim of the present study was to report the metabolic profile of live PA bacteria using high-resolution magic angle spinning (HRMAS) nuclear magnetic resonance spectroscopy (NMR), in combination with 1- and 2-dimensional HRMAS NMR. This methodology provides a new and powerful technique to rapidly interrogate the metabolome of intact bacterial cells and has several advantages over traditional techniques that identify metabolome components from disrupted cells. Furthermore, application of multidimensional HRMAS NMR, in combination with the novel technique total through-Bond correlation Spectroscopy (TOBSY), is a promising approach that may be used to obtain metabolomics information from intact live bacterial cells and can mediate such analyses in a short period of time. Moreover, HRMAS H NMR enables the investigation of the associations between metabolites and cell processes. In the present study, we detected and quantified several informative metabolic molecules in live PA cells, including N-acetyl, betaine, citrulline, alanine and glycine, which are important in peptidoglycan synthesis. The results provided a complete metabolic profile of PA for future studies of PA clinical isolates and mutants. In addition, this NMR biomedical approach might have clinical utility and should prove useful in gene function validation, the study of pathogenetic mechanisms, the classification of microbial strains into functional/clinical groups, the testing of anti-bacterial agents and the determination of metabolic profiles of bacterial mutants.
铜绿假单胞菌（PA）是一种致病性革兰氏阴性菌，广泛存在于自然界，栖息于土壤、水、植物和动物中。PA是人类有害感染的常见病因，尤其是在宿主防御机制受损的患者中。代谢组学是用于研究宿主-病原体相互作用以及鉴定新治疗靶点和相应化合物的重要工具。本研究的目的是使用高分辨率魔角旋转（HRMAS）核磁共振波谱（NMR）结合一维和二维HRMAS NMR报告活PA细菌的代谢谱。该方法提供了一种新的强大技术，可快速探究完整细菌细胞的代谢组，与从破碎细胞中鉴定代谢组成分的传统技术相比具有多个优势。此外，将多维HRMAS NMR与新技术全通过键相关光谱（TOBSY）结合应用，是一种有前景的方法，可用于从完整的活细菌细胞中获取代谢组学信息，并能在短时间内进行此类分析。而且，HRMAS 1H NMR能够研究代谢物与细胞过程之间的关联。在本研究中，我们检测并定量了活PA细胞中的几种信息性代谢分子，包括在肽聚糖合成中起重要作用的N-乙酰基、甜菜碱、瓜氨酸、丙氨酸和甘氨酸。这些结果为PA临床分离株和突变体的未来研究提供了完整的代谢谱。此外，这种NMR生物医学方法可能具有临床实用性，在基因功能验证、致病机制研究、将微生物菌株分类为功能/临床组、抗菌剂测试以及细菌突变体代谢谱测定方面应会证明有用。
