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EspP double mutation L332A/F349A disrupts the structure of the passenger domain locally. AD202 cells transformed with pRLS5 or a derivative encoding the L332A/F349A mutant were subjected to pulse-chase labeling following the addition of IPTG. Radiolabeled cells were removed by centrifugation, and half of the culture medium was treated with PK. Immunoprecipitations were then performed using antisera generated against the EspP passenger domain (A) or an EspP N-terminal peptide (B).
