Comprehensive methodology for Staphylococcus aureus lipidomics by liquid chromatography and quadrupole time-of-flight mass spectrometry

Hewelt-Belka, Weronika; Nakonieczna, Joanna; Belka, Mariusz; Bączek, Tomasz; Namieśnik, Jacek; Kot‐Wasik, Agata

doi:10.1016/j.chroma.2014.08.020

Cited by 33 publications

(39 citation statements)

References 55 publications

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“…The presence of unsaturated lipids in S. aureus is somewhat controversial, with a common assumption in the community that the biophysical role of unsaturated fatty acids is substituted by the abundant branched-chain fatty acids in Gram-positive bacteria [25]. Indeed, in two recent LC-MS studies on S. aureus lipids, one reported only saturated lipids to exist in Col and Newman strains [20], while the other on the N315 strain detected lipids containing unsaturated fatty acid chains (specifically C29:1 to C35:1 PG species) [21], though no comment was made on the existence of these molecular species. However, it is common to have minor variations in fatty acid contents and thus the profiles of bacterial lipidomes due to variations in growth phase, choice of strain and culture media, and indeed these earlier studies utilised brain-heart infusion media [20,21].…”

Section: Resultsmentioning

confidence: 99%

“…Nevertheless, a need exists for a rapid, sensitive and accurate quantitative method to characterise in detail the lipid composition of the S. aureus cell membrane. The recent applications of two distinct liquid chromatographic (LC) MS approaches, reversed-phase LC-MS [20] and hydrophilic interaction LC-ion mobility-MS [21], have addressed some of the limitations in earlier methodologies. Both of these studies describe similar, but not identical S. aureus lipidomes, but these remain incomplete, as the authors did not obtain positional assignments for the individual fatty acid chains to enable a full characterisation of each lipid species.…”

Section: Introductionmentioning

confidence: 99%

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Characterisation ofStaphylococcus aureuslipids by nanoelectrospray ionisation tandem mass spectrometry (nESI-MS/MS)

Young

Desbois

Coote

et al. 2019

Preprint

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19Staphylococcus aureus is a major opportunistic pathogen that is exposed to antimicrobial 20 innate immune effectors and antibiotics that can disrupt its cell membrane. An 21 understanding of S. aureus lipid composition and its role in defending the cell against 22 validated the molecular characterization and showed the principal species present in each 33 strain were predominately anteiso-and iso-branched chain fatty acids. Though the fatty acid 34 and lipid profiles were similar between the S. aureus strains, this method was sufficiently 35 sensitive to distinguish minor differences in lipid composition. In conclusion, this nESI-36 MS/MS methodology can characterise the role of lipids in antimicrobial resistance, and may 37 even be applied to the rapid diagnosis of drug-resistant strains in the clinic. 38 3 39 101 102 6 103 Materials and methods 104 S. aureus culture and lipid extraction 105 All reagents and culture media were purchased from Sigma-Aldrich Ltd. S. aureus Newman 106 (methicillin-susceptible; MSSA), BB270 (methicillin-resistant; MRSA) and Mu50 (MRSA and 107 vancomycin intermediate-resistant) were sourced and cultured in Müller-Hinton broth as 108 described previously [22]. Briefly, each bacterial strain was prepared by inoculation of a 109 colony from an agar plate into 10 ml Müller-Hinton broth and cultured at 37°C with shaking 110 until reaching an OD 600nm = 1 (~1 × 10 9 colony-forming units [cfu]). Bacterial cells were 111 harvested (1000 × g, 10 min), washed in PBS before a repeat centrifugation and removal of 112 the supernatant. Lipids were extracted according to the Bligh-Dyer method [23], dried under 113 nitrogen, and stored at 4°C until analysis by mass spectrometry. 114 115 157

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Characterisation ofStaphylococcus aureuslipids by nanoelectrospray ionisation tandem mass spectrometry (nESI-MS/MS)

Young

Desbois

Coote

et al. 2019

Preprint

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“…Examples include metabolomics, phosphoproteomics and lipidomics, and their development and application have been described elsewhere [76,77,78]. While these methods have not been widely used for antibiotic target elucidation, they may facilitate the identification of bacterial components that participate in key aspects of bacterial physiology that have not yet been explored as potential targets for antibiotic development.…”

Section: Transcriptomics and Proteomics In Antibiotic Target Discomentioning

confidence: 99%

Using bacterial genomes and essential genes for the development of new antibiotics

Fields

Lee

McConnell

2017

Biochemical Pharmacology

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The shrinking antibiotic development pipeline together with the global increase in antibiotic resistant infections requires that new molecules with antimicrobial activity are developed. Traditional empirical screening approaches of natural and non-natural compounds have identified the majority of antibiotics that are currently available, however this approach has produced relatively few new antibiotics over the last few decades. The vast amount of bacterial genome sequence information that has become available since the sequencing of the first bacterial genome more than 20 years ago holds potential for contributing to the discovery of novel antimicrobial compounds. Comparative genomic approaches can identify genes that are highly conserved within and between bacterial species, and thus may represent genes that participate in key bacterial processes. Whole genome mutagenesis studies can also identify genes necessary for bacterial growth and survival under different environmental conditions, making them attractive targets for the development of novel inhibitory compounds. In addition, transcriptomic and proteomic approaches can be used to characterize RNA and protein levels on a cellular scale, providing information on bacterial physiology that can be applied to antibiotic target identification. Finally, bacterial genomes can be mined to identify biosynthetic pathways that produce many intrinsic antimicrobial compounds and peptides. In this review, we provide an overview of past and current efforts aimed at using bacterial genomic data in the discovery and development of novel antibiotics.

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“…Lipidomics, the metabolomic approach targeting lipids by MS is also a promising approach to identify pathogens owing to the huge number of lipids and their important cellular roles. Microbial lipidomics also addresses crucial biological questions such as the understanding of the role of cell wall lipids in the pathology, [162] the comparative analysis of clinical isolates and strains, [163,164] the study of the mechanism of viral replication and host-cell interactions, [165,166] or as biomarkers of infections. [167][168][169][170] Breathomics A specific field of metabolomics is breathomics, which consists of the analysis of volatile organic compounds (VOCs) in the human exhaled breath.…”

Section: Metabolomicsmentioning

confidence: 99%

Mass spectrometry for the detection of bioterrorism agents: from environmental to clinical applications

Duriez¹,

Armengaud

Fenaille

et al. 2016

J. Mass Spectrom.

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In the current context of international conflicts and localized terrorist actions, there is unfortunately a permanent threat of attacks with unconventional warfare agents. Among these, biological agents such as toxins, microorganisms, and viruses deserve particular attention owing to their ease of production and dissemination. Mass spectrometry (MS)-based techniques for the detection and quantification of biological agents have a decisive role to play for countermeasures in a scenario of biological attacks. The application of MS to every field of both organic and macromolecular species has in recent years been revolutionized by the development of soft ionization techniques (MALDI and ESI), and by the continuous development of MS technologies (high resolution, accurate mass HR/AM instruments, novel analyzers, hybrid configurations). New possibilities have emerged for exquisite specific and sensitive detection of biological warfare agents. MS-based strategies for clinical application can now address a wide range of analytical questions mainly including issues related to the complexity of biological samples and their available volume. Multiplexed toxin detection, discovery of new markers through omics approaches, and identification of untargeted microbiological or of novel molecular targets are examples of applications. In this paper, we will present these technological advances along with the novel perspectives offered by omics approaches to clinical detection and follow-up.

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Comprehensive methodology for Staphylococcus aureus lipidomics by liquid chromatography and quadrupole time-of-flight mass spectrometry

Cited by 33 publications

References 55 publications

Characterisation ofStaphylococcus aureuslipids by nanoelectrospray ionisation tandem mass spectrometry (nESI-MS/MS)

Characterisation ofStaphylococcus aureuslipids by nanoelectrospray ionisation tandem mass spectrometry (nESI-MS/MS)

Using bacterial genomes and essential genes for the development of new antibiotics

Mass spectrometry for the detection of bioterrorism agents: from environmental to clinical applications

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