biostudies-literatureRivera SLAmerican Cancer SocietyVetenskapsrådetKnut och Alice Wallenbergs StiftelseNational Institutes of HealthNIGMS NIH HHSKempe FoundationNational Science Foundation213-220.e4https://www.ebi.ac.uk/biostudies/studies/S-EPMC7897265biostudies-literatureUnknown28(2)Transpeptidation reinforces the structure of cell-wall peptidoglycan, an extracellular heteropolymer that protects bacteria from osmotic lysis. The clinical success of transpeptidase-inhibiting β-lactam antibiotics illustrates the essentiality of these cross-linkages for cell-wall integrity, but the presence of multiple, seemingly redundant transpeptidases in many species makes it challenging to determine cross-link function. Here, we present a technique to link peptide strands by chemical rather than enzymatic reaction. We employ biocompatible click chemistry to induce triazole formation between azido- and alkynyl-d-alanine residues that are metabolically installed in the peptidoglycan of Gram-positive or Gram-negative bacteria. Synthetic triazole cross-links can be visualized using azidoCell chemical biologyChemically Induced Cell Wall Stapling in Bacteria.PMC7897265T32 GM120007R25 GM086264Espaillat AShieh PAditham AKRivera SLKim JMuriel-Mundo CCava FSiegrist MSfalseChemically Induced Cell Wall Stapling in Bacteria.Transpeptidation reinforces the structure of cell-wall peptidoglycan, an extracellular heteropolymer that protects bacteria from osmotic lysis. The clinical success of transpeptidase-inhibiting β-lactam antibiotics illustrates the essentiality of these cross-linkages for cell-wall integrity, but the presence of multiple, seemingly redundant transpeptidases in many species makes it challenging to determine cross-link function. Here, we present a technique to link peptide strands by chemical rather than enzymatic reaction. We employ biocompatible click chemistry to induce triazole formation between azido- and alkynyl-d-alanine residues that are metabolically installed in the peptidoglycan of Gram-positive or Gram-negative bacteria. Synthetic triazole cross-links can be visualized using azido2021-01-01T00:00:00Z2021 Feb2025-04-04T02:25:08.968Z2025-04-04T02:25:08.968ZS-EPMC78972653323815810.1016/j.chembiol.2020.11.006