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. Author manuscript; available in PMC: 2008 Aug 23.
Published in final edited form as: Biochemistry. 2006 Jan 10;45(1):272–282. doi: 10.1021/bi052014t

Figure 6.

Figure 6

(A) ESI-MS of the 30 minute reaction of HST2 H135A with 1 mM DTT, 400 μM AcH3 (11-mer), 500 μM NAD+, 50 μM HST2 H135A, 20% v/v MeOH, and 20 mM pyridine buffer at pH 7 at 25 °C. HST2 H135A forms ADPr, 1’-O-methyl-ADPr, and OAADPr under these conditions. (B) ESI-MS of the 30 minute control reaction of HST2 WT with 1 mM DTT, 400 μM AcH3 (11-mer), 500 μM NAD+, 21 μM HST2 WT, 20% v/v MeOH, and 20 mM pyridine buffer at pH 7 at 25 °C. HST2 WT forms no detectable 1’-O-methyl-ADPr under these reaction conditions. (C) ESI-MS of the 30 minute non-enzymatic incorporation of 20% v/v MeOH into 500 μM ADPr with 1 mM DTT, 400 μM AcH3 (11-mer), and 20 mM pyridine buffer at pH 7 at 25 °C. There was no detectable 1’-O-methyl-ADPr in this control. Therefore, the 1’-O-methyl-ADPr observed with HST2 H135A was due to enzymatic activity inherent in the mutant enzyme only.