Fig. 6: De-MARylation of the MERS-CoV macro domain at different temperatures.
From: Elucidating the tunability of binding behavior for the MERS-CoV macro domain with NAD metabolites
A The deMAR enzyme activity of the MERS-CoV macro domain. Auto-mono-ADP-ribosylated human ARTD 10 catalytic domain (HsARTD10-CatD) was arranged using biotin-NAD+ as a substrate. Budding yeast macro domain Poa1p (ScPoa1p) is a positive control. Varying amounts of the MERS-CoV macro domain (from 2.5 to 10.0 μM) mixing with biotin labeled MAR-HsARTD10-CatD at 298 and 308 K were resolved by 15% acrylamide gel. The level of mono-ADP-ribosylation was detected by western blot assay using antibody anti-biotin. B The relative intensity of the bands corresponding to MAR-remained after the reactions of the MERS-CoV macro domain at different temperatures. Independent experiments n = 3. The deMAR enzyme activity of the MERS-CoV macro domain with the presence of C NAD and D ADPR, respectively. Auto-mono-ADP-ribosylated human ARTD 10 catalytic domain (HsARTD10-CatD) was arranged using biotin-NAD+ as a substrate. Varying amounts of C NAD or D ADPR mixing with 20.0 μM MERS-CoV macro domain and biotin-labeled MAR-HsARTD10-CatD at 298 K and 308 K were resolved by 15% acrylamide gel. The level of mono-ADP-ribosylation was detected by western blot assay using antibody antibiotin.
