PimS0 was added in 100 μL reaction mix (10% v/v glycerol, 50 mM NaCl, 20 mM HEPES pH 7.5, 5 mM ATP, 1 mM acetate + CoA/malonate + CoA/acetyl-CoA/malonyl-CoA) in 7.5 mg/mL final concentration to initiate the substrate loading reaction. At 1 h , 4 h, and 24 h, 20 uL reaction solution was quenched with 1% formic acid, flash cooled in liquid N2, and stored at -80 °C for subsequent treatment. PimS0 with no tested substrate added in the reaction mix was taken as a negative control. Proteins in the rea
...ction samples were precipitated by addition of 1 mM NaCl and 4 x vol acetone, followed by two additional washes with 80% acetone in water[Show more]