Abstract
Most bacteria break down a significant portion of their cell wall peptidoglycan during each round of growth and cell division. This process generates peptidoglycan fragments of various sizes that can either be imported back into the cytoplasm for recycling or released from the cell. Released fragments have been shown to act as microbe-associated molecular patterns for the initiation of immune responses, as triggers for the initiation of mutualistic host–microbe relationships, and as signals for cell–cell communication in bacteria. Characterizing these released peptidoglycan fragments can, therefore, be considered an important step in understanding how microbes communicate with other organisms in their environments. In this chapter, we describe methods for labeling cell wall peptidoglycan, calculating the rate at which peptidoglycan is turned over, and collecting released peptidoglycan to determine the abundance and species of released fragments. Methods are described for both the separation of peptidoglycan fragments by size-exclusion chromatography and further detailed analysis by HPLC.
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References
Höltje JV (1998) Growth of the stress-bearing and shape-maintaining murein sacculus of Escherichia coli. Microbiol Mol Biol Rev 62(1):181–203
Typas A, Banzhaf M, Gross CA, Vollmer W (2012) From the regulation of peptidoglycan synthesis to bacterial growth and morphology. Nat Rev Micro 10(2):123–136. doi:10.1038/nrmicro2677
Goodell EW (1985) Recycling of murein by Escherichia coli. J Bacteriol 163(1): 305–310
Reith J, Mayer C (2011) Peptidoglycan turnover and recycling in Gram-positive bacteria. Appl Microbiol Biotechnol 92(1):1–11. doi:10.1007/s00253-011-3486-x
Litzinger S, Duckworth A, Nitzsche K, Risinger C, Wittmann V, Mayer C (2010) Muropeptide rescue in Bacillus subtilis involves sequential hydrolysis by beta-N-acetylglucosaminidase and N-acetylmuramyl-l-alanine amidase. J Bacteriol 192(12):3132–3143. doi:10.1128/JB.01256-09
Cloud-Hansen KA, Peterson SB, Stabb EV, Goldman WE, McFall-Ngai MJ, Handelsman J (2006) Breaching the great wall: peptidoglycan and microbial interactions. Nat Rev Microbiol 4(9):710–716. doi:10.1038/nrmicro1486
Dworkin J (2014) The medium is the message: interspecies and interkingdom signaling by peptidoglycan and related bacterial glycans. Annu Rev Microbiol 68:137–154. doi:10.1146/annurev-micro-091213-112844
Cookson BT, Cho HL, Herwaldt LA, Goldman WE (1989) Biological activities and chemical composition of purified tracheal cytotoxin of Bordetella pertussis. Infect Immun 57(7):2223–2229
Sinha RK, Rosenthal RS (1980) Release of soluble peptidoglycan from growing conococci: demonstration of anhydro-muramyl-containing fragments. Infect Immun 29(3):914–925
Vollmer W, Joris B, Charlier P, Foster S (2008) Bacterial peptidoglycan (murein) hydrolases. FEMS Microbiol Rev 32(2):259–286. doi:10.1111/j.1574-6976.2007.00099.x
Jacobs C, Huang LJ, Bartowsky E, Normark S, Park JT (1994) Bacterial cell wall recycling provides cytosolic muropeptides as effectors for beta-lactamase induction. EMBO J 13(19): 4684–4694
Desmarais SM, de Pedro MA, Cava F, Huang KC (2013) Peptidoglycan at its peaks: how chromatographic analyses can reveal bacterial cell wall structure and assembly. Mol Microbiol 89(1):1–13. doi:10.1111/mmi.12266
Magalhaes JG, Philpott DJ, Nahori MA, Jehanno M, Fritz J, Le Bourhis L, Viala J, Hugot JP, Giovannini M, Bertin J, Lepoivre M, Mengin-Lecreulx D, Sansonetti PJ, Girardin SE (2005) Murine Nod1 but not its human orthologue mediates innate immune detection of tracheal cytotoxin. EMBO Rep 6(12):1201–1207. doi:10.1038/sj.embor.7400552
Veyrier FJ, Williams AH, Mesnage S, Schmitt C, Taha MK, Boneca IG (2013) De-O-acetylation of peptidoglycan regulates glycan chain extension and affects in vivo survival of Neisseria meningitidis. Mol Microbiol 87(5): 1100–1112. doi:10.1111/mmi.12153
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This work was supported by the National Institutes of Health through grants AI097157 and AI099539.
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Schaub, R.E., Lenz, J.D., Dillard, J.P. (2016). Analysis of Peptidoglycan Fragment Release. In: Hong, HJ. (eds) Bacterial Cell Wall Homeostasis. Methods in Molecular Biology, vol 1440. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3676-2_14
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DOI: https://doi.org/10.1007/978-1-4939-3676-2_14
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