Figure EV5. Identification of HybA cleavage site by GlpG.

N‐terminal sequencing of cleaved chromosomally encoded sfCherry‐3xFLAG‐tagged HybA by chromosomally encoded GlpG in S. sonnei in the absence of HybB. Bacteria were grown anaerobically in LB supplemented with 0.5% fumarate, 0.5% glycerol to OD₆₀₀ = 0.5 before lysis. HybA was purified by anti‐FLAG affinity chromatography prior to analysis by SDS–PAGE. Two bands only present in the GlpG (+) sample with molecular weight consistent with cleaved HybA were subject to N‐terminal sequencing and gave the same result.