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. Author manuscript; available in PMC: 2008 Aug 12.
Published in final edited form as: Biochemistry. 2006 Jul 4;45(26):8163–8173. doi: 10.1021/bi060446d

FIGURE 3.

FIGURE 3

The HPLC assay to determine TviB activity. Reaction mixture contained 0.5 μM TviB, 5 mM DTT, 2 mM MgCl2, 0.5 mM NAD+, and 0.25 mM of either UDP-Glc (trace 1), UDP-Gal (trace 2), UDP-GalNAc (3, trace 3), or UDP-GlcNAc (2, trace 4) in 20 mM Tris•HCl buffer (pH 7.5). Incubation with UDP-GlcNAc (2) in the absence of NAD+ was also performed (trace 5). Product formation was detected only in the sample using UDP-GlcNAc (2) as substrate and in the presence of NAD+ (trace 4). The product was confirmed to be UDP-GlcNAcA (4) by NMR. See Experimental Procedures for details. Peaks marked * are the contaminants from NAD+.