Abstract
Bacteria cells are protected from osmotic and environmental stresses by an exoskeleton-like polymeric structure called peptidoglycan (PG) or murein sacculus. This structure is fundamental for bacteria’s viability and thus, the mechanisms underlying cell wall assembly and how it is modulated serve as targets for many of our most successful antibiotics. Therefore, it is now more important than ever to understand the genetics and structural chemistry of the bacterial cell walls in order to find new and effective methods of blocking it for the treatment of disease. In the last decades, liquid chromatography and mass spectrometry have been demonstrated to provide the required resolution and sensitivity to characterize the fine chemical structure of PG. However, the large volume of data sets that can be produced by these instruments today are difficult to handle without a proper data analysis workflow. Here, we present PG-metrics, a chemometric based pipeline that allows fast and easy classification of bacteria according to their muropeptide chromatographic profiles and identification of the subjacent PG chemical variability between e.g. bacterial species, growth conditions and, mutant libraries. The pipeline is successfully validated here using PG samples from different bacterial species and mutants in cell wall proteins. The obtained results clearly demonstrated that PG-metrics pipeline is a valuable bioanalytical tool that can lead us to cell wall classification and biomarker discovery.
Highlights
Bacterial cytoplasmic membrane is surrounded by a net-like polymeric structure named the peptidoglycan (PG) cell wall, which provides protection against environmental stresses, osmotic pressure and, defines bacterial shape
Peptidoglycan is a fundamental structure for the survival of most bacteria and it is a major target of the β-lactams antibiotics
A chemometric-based approach for classifying bacterial peptidoglycan data sets certain bacterial species might exhibit distinct PG chemical structures which can even vary in response to environmental cues [2]
Summary
Bacterial cytoplasmic membrane is surrounded by a net-like polymeric structure named the peptidoglycan (PG) cell wall, which provides protection against environmental stresses, osmotic pressure and, defines bacterial shape. Peptidoglycan canonical composition is based on glycan strands of variable length composed of N-acetylglucosamine-β(1!4)-N-acetyl-muramic acid disaccharide repeats crosslinked by means of short peptide stems [1]. A chemometric-based approach for classifying bacterial peptidoglycan data sets certain bacterial species might exhibit distinct PG chemical structures which can even vary in response to environmental cues [2]. A comprehensive analysis of the PG chemical structure in a large number of species and experimental conditions is fundamental for a better understanding of cell wall biology, in particular its role in signalling and environmental adaptation
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