title: (D)-Amino acid chemical reporters reveal peptidoglycan dynamics of an intracellular pathogen.
abstract: Peptidoglycan (PG) is an essential component of the bacterial cell wall. Although experiments with organisms in vitro have yielded a wealth of information on PG synthesis and maturation, it is unclear how these studies translate to bacteria replicating within host cells. We report a chemical approach for probing PG in vivo via metabolic labeling and bioorthogonal chemistry. A wide variety of bacterial species incorporated azide and alkyne-functionalized d-alanine into their cell walls, which we visualized by covalent reaction with click chemistry probes. The d-alanine analogues were specifically incorporated into nascent PG of the intracellular pathogen Listeria monocytogenes both in vitro and during macrophage infection. Metabolic incorporation of d-alanine derivatives and click chemistry detection constitute a facile, modular platform that facilitates unprecedented spatial and temporal resolution of PG dynamics in vivo.
authors.affiliation: Department of Chemistry, University of California, Berkeley, CA 94720, USA.
authors.forename: M Sloan
authors.initials: MS
authors.lastname: Siegrist
authors.forename: Sarah
authors.initials: S
authors.lastname: Whiteside
authors.forename: John C
authors.initials: JC
authors.lastname: Jewett
authors.forename: Arjun
authors.initials: A
authors.lastname: Aditham
authors.forename: Felipe
authors.initials: F
authors.lastname: Cava
authors.forename: Carolyn R
authors.initials: CR
authors.lastname: Bertozzi
mesh_terms: D000409:Alanine / Q000737:chemistry / Q000378:metabolism*; D057930:Click Chemistry; D008089:Listeria monocytogenes / Q000737:chemistry / Q000166:cytology / Q000378:metabolism*; D008968:Molecular Conformation; D056004:Molecular Dynamics Simulation; D010457:Peptidoglycan / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism*
chemical_list: D010457:Peptidoglycan; D000409:Alanine