Cell lysis was performed as described earlier [13 (link)]. In short, cell were harvested, washed, and resuspended in equal volumes of wet cell pellets, 425–600 μm acid-washed glass beads (G8772, Sigma-Aldrich), and ice-cold lysis buffer (100 mM potassium phosphate buffer, 2 mM magnesium chloride, 5 mM DTT, and Pierce™ Protease Inhibitor Tablets). The cells were disrupted at 4 °C by vortexing 10 times for 30 s with a 30 s cooling step between each vortexing. The beads were removed by centrifugation at 500g for 5 min at 4 °C, and the supernatant transferred to a pre-cooled 1.5 mL tube. The protein concentrations of whole cell lysates were determined by Pierce™ 660 nm Protein Assay.
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