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. 2019:1997:111-120.
doi: 10.1007/978-1-4939-9496-0_8.

Peptidoglycan Composition in Neisseria

Ryan E Schaub  1 , Joseph P Dillard  2
Affiliations

Peptidoglycan Composition in Neisseria

Ryan E Schaub et al. Methods Mol Biol. 2019.

Abstract

The composition of Neisseria peptidoglycan has been of scientific interest for over four decades. Initial investigations focused on discovering the mechanisms causing rising rates of antibiotic resistance in N. gonorrhoeae by determining differences in peptidoglycan composition in penicillin susceptible and resistant strains. The discovery that cytotoxic peptidoglycan fragments are also released by Neisseria furthered the interest in peptidoglycan composition. This method describes the purification, enzymatic degradation, and separation of peptidoglycan fragments by high-performance liquid chromatography (HPLC). It also describes the preparation of samples so that they can be positively identified by mass spectrometry.

Keywords: Cell wall; HPLC; O-Acetylation; Peptidoglycan.

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Figures

Fig. 1:
Fig. 1:
Chromatograms showing the peptidoglycan composition of two strains of N. gonorrhoeae. A comparison of strain FA19 and an MS11 pacA mutant defective in O-acetylation. An asterisk (*) shows the locations where peaks containing acetylated PG fragments would appear in a wild-type strain. Some peaks contain two different peptidoglycan fragments, such as the one containing Penta(OAc) and the one containing crosslinked Tetra-tetra monomers and Tri(Anh). FA19 is known to contain more dipeptide than other strains, but it is otherwise similar to other wild-type strains. Labels indicate disaccharides with two (di), three (tri), four (tetra), or five (penta) amino acids. OAc, O-acetylation; Anh, 1,6-anhydromuramyl moiety.
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