An overview of analytical methods for monitoring bacterial transglycosylation

Abstract

This review highlights the fluorescence and radioactively labeled assays and high-throughput screens for the search for antibiotics targeting bacterial transglycosylation.

Advances and prospects of analytic methods for bacterial transglycosylation and inhibitor discovery

“…61 For paper chromatography, isobutyric acid/1 M ammonia solution (5 : 3, v/v) is typically used as the mobile phase. 11 Lipid II bearing long isoprenoid lipid is more hydrophobic than peptidoglycan, and thus less retained on paper chromatography, showing higher R f value (Fig. 4A).…”

“…5 Studies of PG morphology and in vivo imaging provide useful information on the construction of the cell wall and its distribution. [6][7][8][9][10] Current analytic methods used to evaluate TGase activities before 2016 have been covered by several outstanding reviews; [11][12][13][14] however, many new chemistries have since been developed, including those focusing on shape, elongation, division and sporulation (SEDS) proteins. In this review, we shall discuss the up-todate analytic methods for evaluation of the transglycosylation (TG) reaction in cell wall formation.…”

Breaking down the cell wall: Strategies for antibiotic discovery targeting bacterial transpeptidases

“…We then highlight promising recent approaches for assaying the transpeptidase activities of PBP and Ldts, based principally on interactions with substrate analogues and β-lactam-based probes. This review does not cover PBP transglycosylase activity or the sortase family of transpeptidases, which were covered in excellent recent reviews [19][20][21].…”

Development of a liquid chromatography/mass spectrometry assay for the bacterial transglycosylation reaction through measurement of Lipid II

“…The absence of a UV‐chromophore in the structure of Lipid II, led to the development of assays using fluorescent or radiochemical labeling (see for a review). However, label‐free assays are interesting as well because there is no chance of altered affinity.…”

“…The peptide moieties in S. aureus peptidoglycan are cross-linked by a flexible pentaglycine chain allowing a cross-linking degree of 90% yielding a very rigid cell wall structure [13,14]. The absence of a UV-chromophore in the structure of Lipid II, led to the development of assays using fluorescent or radiochemical labeling (see [15] for a review). However, label-free assays are interesting as well because there is no chance of altered affinity.…”