Extended Data Fig. 10: Cleavage of probe sequon variants co-expressed with OGO-6 by ZmpB mutants.
From: A high-throughput screening platform for enzymes active on mucin-type O-glycoproteins
ZmpB mutants were incubated with 2OGO-6 and analyzed at (a) 1.5 hours and (b) 8 hours via SDS-PAGE and in-gel fluorescence, or 4OGO-6 before analysis again at (c) 1.5 hours and (d) 8 hours. Panels e and f show the reaction time course as monitored by densitometry for M6 (white data points) against (e) 2OGO-6 and (f) 4OGO-6. Data from the WT (black data points) is also included to serve as a reference. Note that for these reactions a high concentration of probe (2.5 mg/mL) and ZmpB (0.5 mg/mL) was used in order to better enable visualization of the cleavage products. Gels shown have been overexposed to better show detail and were not used for densitometry calculation. Data points shown are technical replicates.
