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. 2016 Jun 1;291(32):16740–16752. doi: 10.1074/jbc.M116.717827

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© 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 4. — NR consumption of 20 μm (10bS,4aR or 10bR,4aS)-noroxomaritidine using the same assay conditions as for specific activity assays incubated 2 h. A, LC-MS using a Chrom Tech, Inc. Chiral-CBH 100 × 4.0-mm, 5-μm column and the same LC setup and time program as for chiral separations in Ref. 18. Samples were monitored at m/z 272.3 with CE (25) and DP (70) on the QTRAP 6500 for the following samples top to bottom: (10bS,4aR and 10bR,4aS)-noroxomaritidine mixed standard; complete assay with NR, (10bS,4aR and 10bR,4aS)-noroxomaritidine and NADPH; assay without NADPH; assay without (10bS,4aR and 10bR,4aS)-noroxomaritidine substrate; assay without NR enzyme; and assay with TALON resin purified E. coli empty vector protein extract substituted for NR protein. B, the two enantiomers of noroxomaritidine and corresponding expected products.