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Comparison of Bioorthogonal β‐Lactone Activity‐Based Probes for Selective Labeling of Penicillin‐Binding Proteins
Author(s) -
Brown Nathaniel W.,
Shirley Joshua D.,
Marshall Andrew P.,
Carlson Erin E.
Publication year - 2021
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.202000556
Subject(s) - bioorthogonal chemistry , biochemistry , penicillin binding proteins , chemistry , gene isoform , penicillin , antibiotics , combinatorial chemistry , click chemistry , gene
Penicillin‐binding proteins (PBPs) are a family of bacterial enzymes that are key components of cell‐wall biosynthesis and the target of β‐lactam antibiotics. Most microbial pathogens contain multiple structurally homologous PBP isoforms, making it difficult to target individual PBPs. To study the roles and regulation of specific PBP isoforms, a panel of bioorthogonal β‐lactone probes was synthesized and compared. Fluorescent labeling confirmed selectivity, and PBPs were selectively enriched from Streptococcus pneumoniae lysates. Comparisons between fluorescent labeling of probes revealed that the accessibility of bioorthogonal reporter molecules to the bound probe in the native protein environment exerts a more significant effect on labeling intensity than the bioorthogonal reaction used, observations that are likely applicable beyond this class of probes or proteins. Selective, bioorthogonal activity‐based probes for PBPs will facilitate the activity‐based determination of the roles and regulation of specific PBP isoforms, a key gap in knowledge that has yet to be filled.