Figure 3.
Reversed-phase HPLC of radiolabelled fragments. Pooled fractions from size-exclusion chromatography of wild-type N. gonorrhoeae supernatant were analyzed by HPLC with a C18 column. (A) Monomer fractions were run using a 4 to 13% gradient of acetonitrile with 0.05% TFA at 1 mL/min for 30 min. Peaks with retention times of 18 minutes and 22 minutes correspond to 1,6-anhydrodissacharide-tripeptide and -tetrapeptide respectively. Quantification of peaks reveals a 3:1 ratio of tripeptide to tetrapeptide peptidoglycan monomers released. (B) Dimer fractions were run over a 5 to 25% gradient of acetonitrile with 0.05% TFA at 1 mL/min for 60 min. Peaks with retention times of 25 minutes and 31 minutes correspond to glycosidically-linked and peptide-linked dimers respectively.