LAUSR

Abstract Porcine epidemic diarrhea virus (PEDV) is a coronavirus (CoV) discovered in the 1970s that infects the intestinal tract of pigs, resulting in diarrhea and vomiting. It can cause extreme dehydration and death in neonatal piglets. In Asia, modified live attenuated vaccines have been used to control PEDV infection in recent years. However, a new strain of PEDV that belongs to genogroup 2a appeared in the USA in 2013 and then quickly spread to Canada and Mexico as well as Asian and European countries. Due to the less effective protective immunity provided by the vaccines against this new strain, it has caused considerable agricultural and economic loss worldwide. The emergence of this new strain increases the importance of understanding PEDV as well as strategies for inhibiting it. Coronaviral proteases, including main proteases and papain‐like proteases, are ideal antiviral targets because of their essential roles in viral maturation. Here we provide a first description of the expression, purification and structural characteristics of recombinant PEDV papain‐like protease 2, moreover present our finding that 6‐thioguanine, a chemotherapeutic drug, in contrast to its competitive inhibition on SARS‐ and MERS‐CoV papain‐like proteases, is a noncompetitive inhibitor of PEDV papain‐like protease 2. HighlightsPEDV PL2pro exhibits much higher DUB activity than that of other PLpros in spite of their structural similarities.In contrast to its competitive inhibition on SARS‐ and MERS‐CoV PLpros, 6‐thioguanine inhibits PEDV PL2pro allosterically.Putative 6‐thioguanine binding site is proposed to render the blocking loop less flexible and therefore disfavor catalysis.6‐thioguanine can be a lead compound for anti‐coronaviral drug development.