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#cell wall synthesis

2 public questions tagged with this topic.

Which antibiotic binds to peptidoglycan precursors, inhibiting transpeptidation?

Glycopeptide antibiotic activity depends on extracellular substrate capture rather than direct inhibition of enzyme active site, an unusual mechanism among cell wall agents. During late-stage peptidoglycan assembly, the membrane-anchored disaccharide-pentapeptide Lipid II carrying a terminal D-Ala-D-Ala sequence at the stem terminus is the natural recognition motif for penicillin-binding proteins that perform transpeptidation cross-linking. Vancomycin forms a sophisticated network of five hydrogen bonds with carbonyl oxygens and amide nitrogens of this D-Ala-D-Ala dipeptide, creating a large cup-shaped hydrophobic pocket complex that physically shields the scissile bond from enzymatic approach. Consequently both transglycosylation reaction that elongates glycan strands and transpeptidation reaction that cross-links stem peptides are blocked, as glycosyltransferases and transpeptidases cannot access their natural Lipid II substrate. The net effect is accumulation of uncross-linked precursors, futile cycling, weakening of nascent septal wall and activation of endogenous autolysins such as LytA leading to osmotic lysis. Rifampin inhibits RNA polymerase, tetracycline and gentamicin block bacterial ribosomes, so precursor sequestration uniquely describes vancomycin mechanism distinguishing it from beta-lactams that covalently modify PBPs.

Ref: NCBI Bookshelf, Molecular Biology of the Cell, Section: Vancomycin and Lipid II D-Ala-D-Ala Binding.

β-lactam antibiotics inhibit bacterial growth by:

Peptidoglycan assembly requires synthesis of soluble UDP-N-acetylmuramyl pentapeptide in cytoplasm, attachment to undecaprenyl phosphate carrier to form Lipid II, flipping to external leaflet, polymerization by transglycosylases and final cross-linking of pentapeptide stems by transpeptidases called penicillin-binding proteins possessing active-site serine. The transpeptidase recognizes the terminal D-Ala-D-Ala motif and catalyzes cleavage of the terminal D-alanine with concomitant formation of a new peptide bond between meso-diaminopimelate or lysine and D-alanine from adjacent strand, conferring mechanical rigidity essential to resist turgor. Beta-lactam antibiotics including penicillins, cephalosporins, carbapenems are structural mimics of D-Ala-D-Ala dipeptide geometry. The strained four-membered beta-lactam ring acts as a suicide substrate, rapidly acylating the active-site serine of PBPs and forming a stable, long-lived acyl-enzyme intermediate that is hydrolyzed extremely slowly. Transpeptidation cannot proceed, new cross-links fail to form, and housekeeping autolysins continue to remodel old wall causing net wall degradation. Cells swell, lose shape and lyse, explaining activity preferentially against actively growing cells, Gram-positive susceptibility and resistance via beta-lactamase production or PBP2a expression.

Ref: Lodish et al., Molecular Cell Biology, 8th ed., Chapter 17: Peptidoglycan Synthesis and Beta-Lactam Action.