Use of the fused NS4A peptide-NS3 protease domain to study the importance of the helicase domain for protease inhibitor binding to hepatitis C virus NS3-NS4A

Biochemistry
Diane ThibeaultPeter D White

Abstract

The NS3 protein of hepatitis C virus is unusual because it encodes two unrelated enzymatic activities in linked protease and helicase domains. It has also been intensively studied because inhibitors targeting its protease domain have potential to significantly improve treatment options for those infected with this virus. Many enzymological studies and inhibitor discovery programs have been carried out using the isolated protease domain in complex with a peptide derived from NS4A which stimulates activity. However, some recent publications have suggested that the NS3 helicase domain may influence inhibitor binding and thus suggest work should focus on the full-length NS3-NS4A protein. Here we present the characterization of a single-chain protease in which the NS4A peptide activator is linked to the N-terminus of the NS3 protease domain. This protein behaves well in solution, and its protease activity is very similar to that of full-length NS3-NS4A. We find that this fusion protein, as well as the noncovalent complex of the NS4A peptide with NS3, gives similar Ki values, spanning 3 orders of magnitude, for a set of 25 structurally diverse inhibitors. We also show that simultaneous mutation of three residues on the surface of the...Continue Reading

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Citations

Dec 14, 2011·Proceedings of the National Academy of Sciences of the United States of America·Nikolaus SchieringUrsula Bodendorf
Dec 29, 2012·The Journal of Biological Chemistry·Jeff A O'MearaMontse Llinàs-Brunet
Aug 13, 2013·PloS One·Olga AbianAdrian Velazquez-Campoy
Jan 23, 2014·Journal of Medicinal Chemistry·Åsa RosenquistMaria Beumont-Mauviel
Jan 29, 2014·Journal of Medicinal Chemistry·Steven R LaPlanteSteven J Taylor
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Sep 8, 2011·Current Protocols in Pharmacology·Brian SchultzWilliam E Delaney
Jun 20, 2013·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Amandine Verga-GérardLaure Perrin-Cocon

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