Abstract
Bacterial cells are encompassed by a macromolecular network of peptidoglycan, the major component of bacterial cell walls that provides the cell wall strength and determines bacterial morphology. Eukaryotic organisms sense the presence of bacteria by detecting peptidoglycan fragments shed by the bacterium or released through hydrolytic attack by host enzymes. As such, peptidoglycan plays a major role in regulation of host inflammatory homeostasis and has a remarkable variety of influences on the physiological development of the host.
This chapter describes the tools necessary to characterize both the biochemistry and biological activity of peptidoglycans purified from a broad range of bacteria. These protocols detail the procedures for purification of peptidoglycan and analysis of its composition by reversed-phase high-performance liquid chromatography (HPLC), taking into account the specific requirements for different types of bacteria, and describe a NF-κB-luciferase reporter assay performed using the HEK293T human epithelial cell line, to assess the biological activity of purified peptidoglycans through recognition by mammalian Nod-like receptors Nod1 and Nod2.
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© 2015 Springer Japan
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Wheeler, R., Veyrier, F., Werts, C., Boneca, I.G. (2015). Peptidoglycan and Nod Receptor. In: Taniguchi, N., Endo, T., Hart, G., Seeberger, P., Wong, CH. (eds) Glycoscience: Biology and Medicine. Springer, Tokyo. https://doi.org/10.1007/978-4-431-54841-6_147
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DOI: https://doi.org/10.1007/978-4-431-54841-6_147
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Publisher Name: Springer, Tokyo
Print ISBN: 978-4-431-54840-9
Online ISBN: 978-4-431-54841-6
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