Author: Qi Liu; Amita Gupta; Ayse Okesli-Armlovich; Wenjie Qiao; Curt R. Fischer; Mark Smith; Jan E. Carette; Michael C. Bassik; Chaitan Khosla
Title: Enhancing the Antiviral Efficacy of RNA-Dependent RNA Polymerase Inhibition by Combination with Modulators of Pyrimidine Metabolism Document date: 2020_3_25
ID: 1zk64gsg_12
Snippet: In order to guide our SAR studies, we developed an enzymatic assay to evaluate the inhibitory effect of each CPU analog against recombinant human UCK2, which was expressed and purified in E. coli. A continuous assay system was optimized by coupling UCK2 activity to the pyruvate kinase (PK) reaction, which in turn was coupled to lactate dehydrogenase (LDH) (Tomoike et al., 2017) . Overall reaction progress was continuously monitored by detecting t.....
Document: In order to guide our SAR studies, we developed an enzymatic assay to evaluate the inhibitory effect of each CPU analog against recombinant human UCK2, which was expressed and purified in E. coli. A continuous assay system was optimized by coupling UCK2 activity to the pyruvate kinase (PK) reaction, which in turn was coupled to lactate dehydrogenase (LDH) (Tomoike et al., 2017) . Overall reaction progress was continuously monitored by detecting the UV absorption change at 340 nm, which was correlated with the ATP consumption by UCK2.
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