Extended Data Fig. 1: Designed minimal precursor recapitulates the reactivity of PsnB.
From: Molecular mechanism underlying substrate recognition of the peptide macrocyclase PsnB
a, Sequence logo of precursors of Group 2a graspetides including the leader peptide (red region) and one core motif (blue region). The LFIEDL region is highly conserved in Group 2a graspetides (yellow box). b,c, ATPase assays titrating with ATP (b) in solutions containing 0.4 μM PsnB, 200 μM minimal precursor (MP), 100 mM Tris pH 7.3, 50 mM KCl, and 10 mM MgCl2, or titrating with MgCl2 (c) in solutions containing 0.4 μM PsnB, 200 μM MP, 100 mM Tris pH 7.3, 50 mM KCl, and 5 mM ATP. Data are presented as dot plots with mean ± 1 SD (n = 3 independent experiments) and fitted to a hyperbolic equation. d, Minimal precursor was successfully modified by PsnB with a modification rate similar to that of wild-type PsnA2. 50 μM MP (top panel; the product contains two ester bonds) or wild-type PsnA2 (bottom panel; the product contains eight ester bonds) was co-incubated with 0.5 μM PsnB in buffer A (100 mM Tris pH 7.3, 50 mM KCl, 5 mM ATP, and 10 mM MgCl2) at 37 °C, and the reaction solutions at designated time points were analyzed by MALDI-MS.
