Herod, MR;
Prince, CA;
Skilton, RJ;
Ward, VK;
Cooper, JB;
Clarke, IN;
(2014)
Structure-based design and functional studies of novel noroviral 3C protease chimaeras offer insights into substrate specificity.
Biochemical Journal
, 464
(3)
pp. 461-472.
10.1042/BJ20140959.
Preview |
Text
Structure-based design and functional studies of novel noroviral 3C protease chimaeras offer insights into substrate specificity.pdf Download (969kB) | Preview |
Abstract
The norovirus NS6 protease is a key target for anti-viral drug development. Noroviruses encode a 2200 amino acid polyprotein which is cleaved by this critical protease at five defined boundary substrates into six mature non-structural (NS) proteins. Studies of the human norovirus (HNV) NS6 protease, in the context of a full ORF1 polyprotein, have been severely hampered because HNVs are not culturable. Thus, investigations into the HNV NS6 protease have been largely restricted to in vitro assays using Escherichia coli-expressed, purified enzyme. The NS6 protease is formed of two distinct domains joined by a linking loop. Structural data suggest that domain 2 of the protease possesses substantial substrate binding pockets which form the bulk of the interactions with the NS boundaries and largely dictate boundary specificity and cleavage. We have constructed chimaeric murine norovirus (MNV) genomes carrying individual domains from the HNV protease and demonstrated by cell transfection that chimaeric HNV proteases have functional activity in the context of the full-length ORF1 polyprotein. Although domain 2 primarily confers boundary specificity, our data suggest that an inter-domain interaction exists within HNV NS6 protease which influences cleavage of specific substrates. The present study also shows that chimaeric MNVs provide improved models for studying HNV protein function in the context of a full ORF1 polyprotein.
| Type: | Article |
|---|---|
| Title: | Structure-based design and functional studies of novel noroviral 3C protease chimaeras offer insights into substrate specificity |
| Location: | England |
| Open access status: | An open access version is available from UCL Discovery |
| DOI: | 10.1042/BJ20140959 |
| Publisher version: | http://dx.doi.org/10.1042/BJ20140959 |
| Language: | English |
| Additional information: | Copyright © 2014 The Authors. This is an Open Access article distributed under the terms of the Creative Commons Attribution Licence (CC-BY) (http://creativecommons.org/licenses/by/3.0/) which permits unrestricted use, distribution and reproduction in any medium, provided the original work is properly cited. |
| Keywords: | Amino Acid Sequence, Binding Sites, Crystallography, X-Ray, Cysteine Endopeptidases, HEK293 Cells, Humans, Models, Molecular, Molecular Structure, Norovirus, Proteolysis, Recombinant Fusion Proteins, Substrate Specificity, Viral Proteins |
| UCL classification: | UCL UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences > Div of Medicine |
| URI: | https://discovery-pp.ucl.ac.uk/id/eprint/1458522 |
Archive Staff Only
 |
View Item |
