Extended Data Fig. 1: Further evaluation data for Brc3DPQ1 covalent tethering with the preQ1 riboswitch.
From: Engineering covalent small molecule–RNA complexes in living cells
a, Relative conversion–time plot at different pH for the reaction between Brc3DPQ1 and C15U preQ1 RNA (conditions: 2.5 μM RNA, 60 μM Brc3DPQ1, 100 mM KCl, 2.0 mM MgCl2, 50 mM buffer, 37 °C, pH values as indicated). Mean values (filled circles) ± s.e.m. are shown (n = 3). b, Time course of the reaction between Brc3DPQ1 and Tt preQ1 RNA (conditions as for a) followed by AE HPLC analysis at the time points indicated; conditions as in a but 125 μM Brc3DPQ1. c, Brc3DPQ1 stability at pH 7.5 (50 mM MOPS) at time points 0 and 24 h analyzed by LC-MS experiments. The major degradation product is consistent with intramolecular cyclization; Rp-HPLC (XBridge BEH C18 Column): 0-15% acetonitrile in aqueous TFA (0.1%), in 15 min. d, Brc3DPQ1 stability at pH 6.0 and pH 7.5, respectively. Mean values (filled circles) ± s.e.m. are shown (n = 3).
