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. 2010 Oct 27;285(53):41232–41243. doi: 10.1074/jbc.M110.156273

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FIGURE 5. — Determination of CwlP-M23 cleavage sites using RP-HPLC. B. subtilis peptidoglycan (containing no anionic polymers) was cleaved without (panel A) or with (panel B) CwlP-M23 for 30 min. The free amino acid residues in the samples were labeled with 1-fluoro-2,4-dinitrobenzene and the samples were hydrolyzed with HCl. Samples were separated by RP-HPLC. Both samples contained only the material derived from peak 1 (mono-DNP-A₂pm) or peak 2 (DNP). An arrow indicates the (54 min) retention time of bis-DNP-A₂pm. The peptidoglycan sample digested with CwlP-M23 and LytF (dl-endopeptidase) gave three peaks; peaks 1 and 2, and a new peak derived from bis-DNP-A₂pm that had a 54-min retention time on RP-HPLC (supplemental Fig. S4).