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Mycobacterium tuberculosis ESAT‐6 (MtbESAT‐6) plays essential roles in pathogenesis. MtbESAT‐6 exhibits a unique pore‐forming activity (PFA) that is not found in its ortholog from non‐pathogenic Mycobacterium smegmatis (MsESAT‐6). Here, we characterized the differential PFAs and found that exchange of I25‐H26/T25‐A26 between two proteins reciprocally affected their PFAs. MtbESAT‐6(IH/TA) had ~ 40%...
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