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. Author manuscript; available in PMC: 2020 Apr 23.
Published in final edited form as: Angew Chem Int Ed Engl. 2019 Apr 1;58(18):5962–5966. doi: 10.1002/anie.201900768

Figure 5.

Figure 5.

Heavy NSH serves as a substrate in a non-radioactive 2D 1H-13C HSQC NMR epimerase assay. a) Site-specific 1H incorporation at hexuronic acid C5 positions along the heavy NSH chain is achieved by enzymatic GlcA epimerization and subsequent interconversion of GlcA and IdoA residues, reactions involving C5 2H abstraction and replacement by 1H from solvent (H2O). b) Integration of GlcA (C5,H5) peak indicates incorporation of 1H at GlcA C5 following C5 epimerase treatment. c) 1H-13C HSQC spectra of heavy NSH before (left) and after (right) incorporation of 1H at C5 of hexuronic acid by epimerase treatment in H2O.