Expression, Purification and Characterization of a GII.4 Norovirus Protease from Minerva Virus

Abstract

Compound A, a potent inhibitor of MVpro, is a good starting point for the design of inhibitors to target GII.4 noroviruses. Furthermore, the results presented here will allow for future characterization of MVpro inhibitors as they are synthesized.

Structural and Antiviral Studies of the Human Norovirus GII.4 Protease

“…Initially, expression and purification of the iNoV GII.4 PR revealed the presence of both a monomer and dimer in solution. 34 Crystallographic studies support the formation of the protease dimer in solution. As described previously, norovirus GII.4 protease consists of eight molecules per asymmetric unit subsequently composed of four asymmetric dimers.…”

“…We report here the elucidation of the X-ray crystallographic structure, computational analyses, and the enzymatic inhibition of norovirus GII.4 minerva virus protease. Initially, expression and purification of the iNoV GII.4 PR revealed the presence of both a monomer and dimer in solution . Crystallographic studies support the formation of the protease dimer in solution.…”

“…Compound IC 50 experiments were designed based on enzymatic activity previously described. 34 GII.4 minerva virus protease and GI.1 Norwalk virus protease were diluted in buffer (50 mM HEPES pH 8.0, 120 mM NaCl, 0.4 mM EDTA, 20% glycerol (v/v), 4 mM DTT) to a final well concentration of 128 nM and incubated at 37 °C for 10 min. Compounds A and B were serially diluted in DMSO for a final well concentration of 64 μM-62.5 nM and incubated with diluted protein for 90 min at 37 °C.…”