Antibiotic resistance is rapidly becoming a global healthcare crisis. Bacteria have evolved clever strategies to escape the toxic effects of antibiotics at the same time that the number of FDA antibiotic approvals has dwindled. The discovery of new drug targets and re-evaluation of established targets will be critical in reversing this trend. Potent antibiotics (e.g. vancomycin) target the bacterial cell wall by arresting key intermediates in the cell wall biosynthetic pathway. We hypothesized that installation of chemically modified intermediates in the bacterial cell wall biosynthetic pathway could map out key enzymatic interactions that are crucial for the generation of the bacterial cell wall and thus provide interesting targets for the development of novel antibiotics. Therefore we synthesized photo-affinity based cell wall precursor analogues as metabolic probes and incorporated them in live bacterial cells by hijacking the cell wall biosynthetic pathway. Photo-irradiation in live bacterial cells followed by activity based protein profiling (ABPP) afforded a detailed map of several key enzymes that are supposed to be involved in the biosynthesis of the bacterial cell wall. Out of a total of ~900 proteins identified, ~600 proteins were found to be more abundant in the presence of light. We anticipate that our strategy will be an integral component in revealing the full picture of enzymatic interactions along the bacterial cell wall biosynthetic pathway and can be used as a model for the development of next generation antibiotics.

Mapping enzymatic interactions of bacterial cell wall biosynthesis with photo-affinity probes

251st National Meeting (2016)

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