Extended Data Fig. 5: Bioinformatic identification and biochemical characterization of phage SBSphiJ gene 147 as the anti-Pycsar nuclease Apyc1.
From: Phage anti-CBASS and anti-Pycsar nucleases subvert bacterial immunity
a, Genome schematic of SBSphiJ and 7 other closely related phages highlighting regions exclusive to cCMP-cleaving phages. b, Summary of HHpred analysis of candidate genes. c, Recombinant SBSphiJ Apyc1 was expressed as an N-terminal 6×His-SUMO2 fusion, purified by Ni-NTA, and separated from His-SUMO2 by size exclusion chromatography. d, Coomassie-stained SDS-PAGE analysis of fully purified SBSphiJ Apyc1. e, TLC analysis of cCMP degradation by SBSphiJ Apyc1 at the indicated pH. f, HPLC analysis of recombinant SBSphiJ Apyc1 incubated with the indicated substrates (100 μM) for 30 min at 37 °C. Data in c–f are representative of 3 independent experiments.
