title: In Vivo Probe of Lipid II-Interacting Proteins.
abstract: β-Lactams represent one of the most important classes of antibiotics discovered to date. These agents block Lipid II processing and cell wall biosynthesis through inactivation of penicillin-binding proteins (PBPs). PBPs enzymatically load cell wall building blocks from Lipid II carrier molecules onto the growing cell wall scaffold during growth and division. Lipid II, a bottleneck in cell wall biosynthesis, is the target of some of the most potent antibiotics in clinical use. Despite the immense therapeutic value of this biosynthetic pathway, the PBP-Lipid II association has not been established in live cells. To determine this key interaction, we designed an unnatural d-amino acid dipeptide that is metabolically incorporated into Lipid II molecules. By hijacking the peptidoglycan biosynthetic machinery, photoaffinity probes were installed in combination with click partners within Lipid II, thereby allowing, for the first time, demonstration of PBP interactions in vivo with Lipid II.
authors.affiliation: Department of Chemistry, Lehigh University, Bethlehem, PA, 18015, USA.
authors.forename: Sourav
authors.initials: S
authors.lastname: Sarkar
authors.affiliation: Department of Microbiology & Immunology, Columbia University, New York, NY, 10032, USA.
authors.forename: Elizabeth A
authors.initials: EA
authors.lastname: Libby
authors.affiliation: Department of Chemistry, Lehigh University, Bethlehem, PA, 18015, USA.
authors.forename: Sean E
authors.initials: SE
authors.lastname: Pidgeon
authors.affiliation: Department of Microbiology & Immunology, Columbia University, New York, NY, 10032, USA.
authors.forename: Jonathan
authors.initials: J
authors.lastname: Dworkin
authors.affiliation: Department of Chemistry, Lehigh University, Bethlehem, PA, 18015, USA. map311@lehigh.edu.
authors.forename: Marcos M
authors.initials: MM
authors.lastname: Pires
mesh_terms: D001412:Bacillus subtilis / Q000166:cytology / Q000378:metabolism; D002473:Cell Wall / Q000378:metabolism; D015394:Molecular Structure; D046915:Penicillin-Binding Proteins / Q000737:chemistry* / Q000378:metabolism; D014538:Uridine Diphosphate N-Acetylmuramic Acid / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism
chemical_list: D046915:Penicillin-Binding Proteins; D014538:Uridine Diphosphate N-Acetylmuramic Acid; C069249:muramyl-NAc-(pentapeptide)pyrophosphoryl-undecaprenol
keywords: amino acids; cell walls; lipid II; peptides; proteomics