1887

Abstract

A recombinant form of yellow fever virus (YFV) NS3 protease, linked via a nonapeptide to the minimal NS2B co-factor sequence (CF40-gly-NS3pro190), was expressed in and shown to be catalytically active. It efficiently cleaved the fluorogenic tetrapeptide substrate Bz-norleucine-lysine-arginine-arginine-AMC, which was previously optimized for dengue virus NS2B/3 protease. A series of small peptidic inhibitors based on this substrate sequence readily inhibited its enzymic activity. To understand the structure–activity relationship of the inhibitors, they were docked into a homology model of the YFV NS2B/NS3 protease structure. The results revealed that the P1 and P2 positions are most important for inhibitor binding, whilst the P3 and P4 positions have much less effect. These findings indicate that the characteristics of YFV protease are very similar to those reported for dengue and West Nile virus proteases, and suggest that pan-flavivirus NS3 protease drugs may be developed for flaviviral diseases.

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2007-08-01
2019-10-17
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Supplements

vol. , part 8, pp. 2223 – 2227

Expression of recombinant YFV CF40-gly-NS3pro190 protein

IC determination and active-site titration

Expression, purification and activity test of mutant YFV CF40-gly-NS3pro190 (S138A) protease

Enzymic characterization of YFV CF40-gly-NS3pro190

Generation of wild-type and S138A mutant YFV CF40-gly-NS3pro190 expression constructs and homology modelling

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