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. 2022 Feb 22;119(8):e2110293119. doi: 10.1073/pnas.2110293119

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Fig. 8. — Release of zinc from kupyaphores is mediated by Mtb isonitrile hydratase. (A) Reaction scheme for in vitro enzymatic assay of Rv0052. (B) MS/MS spectra with fragmentation pattern and products marked for expected product 3, confirming the presence of formamide product in the assay. The observed masses are within 10 ppm mass error tolerance at MS1 or MS/MS for each mass. (C) qRT based mRNA expression analysis of Rv0052 in WT Mtb under low- and high-zinc condition as compared to optimal-zinc growth condition. (D) Reaction scheme for Rv0052 enzymatic assay with kupyaphores. (E) Relative MS intensities of kupyaphore species VI of m/z for [M+H]⁺= 848.69, and corresponding monoformamide 4 and diformamide 5 analogs in WT Mtb extract treated with or without Rv0052. The calculated masses for metabolites are within 5 ppm mass error tolerance at MS1. All assays in B and E were performed in triplicates and duplicates, respectively, with reproducible results each time.