Extended Data Fig. 5: Characterization of V273A and W84K mutations.
From: Bacterial TIR-based immune systems sense phage capsids to initiate defense
(a) Structure Φ80α Mhp monomer (PDB: 6B0X), colored according to the different structural domains (red, N-arm; yellow, P domain and loop; green, A-domain; orange, E-loop). (b) Structure of the hexameric form of Φ80α Mhp (PDB: 6B0X), with one capsomer interface highlighted. The position of the residues which mutations that led to immune evasion, V273 within the P-loop (tan) and W84 within the E-loop (brown), are shown (ipTM 0.47, pTM 0.52). (c) Coomassie Blue-stained SDS-PAGE of proteins purified from S. aureus expressing His6-ThsA (56.0 kDa) using a cobalt resin. Protein molecular weight (kDa) markers are shown. (d) Coomassie Blue-stained SDS-PAGE of Mhp proteins, wild type and V273A mutant, purified from staphylococci harboring pMhp plasmids, using PEG-enrichment. Protein molecular weight (kDa) markers are shown. (e) Hydrolysis reaction of oxidized nicotinamide adenine dinucleotide (NAD + ) mediated by ThsA when activated by 1”-3’-gcADPR, which yields ADP-ribose (ADPR) and nicotinamide (NAM).
