Features of QTF, a fluorogenic probe of Ag85-mediated cell wall biogenesis. (A) Components of the mycobacterial cell envelope include the peptidoglycan-anchored mycolyl-arabinogalactan (mAG) complex. The mycolyltransferases, including the antigen 85 complex (Ag85) of M. tuberculosis, construct the mycolic acid membrane by processing endogenous trehalose monomycolate (TMM) to afford trehalose dimycolate (TDM) or mycolyl-arabinogalactan (mAG). (B) QTF is a TMM mimic bearing a fluorophore and quencher. QTF processing by mycolyltransferases generates a fluorescent signal. (C) Chemical structure of TMM, QTF, and QTF products Q-Tre and lipid-FL generated by Ag85 cleavage.