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Structure-guided mutagenesis of active site residues in the dengue virus two-component protease NS2B-NS3 by Wanisa Salaemae; Muhammad Junaid; Chanan Angsuthanasombat; Gerd Katzenmeier is a Medicine article available to read on EtoBox.

What is Structure-guided mutagenesis of active site residues in the dengue virus two-component protease NS2B-NS3 about?

## Abstract ## Background The dengue virus two-component protease NS2B/NS3 mediates processing of the viral polyprotein precursor and is therefore an important determinant of virus replication. The enzyme is now intensively studied with a view to the structure-based development of antiviral inhibitors. Although 3-dimensional structures have now been elucidated for a number of flaviviral proteases, enzyme-substrate interactions are characterized only to a limited extend. The high selectivity of the dengue virus protease for the polyprotein precursor offers the distinct advantage of designing inhibitors with exquisite specificity for the viral enzyme. To identify important determinants of substrate binding and catalysis in the active site of the dengue virus NS3 protease, nine residues, L115, D129, G133, T134, Y150, G151, N152, S163 and I165, located within the S1 and S2 pockets of the enzyme were targeted by alanine substitution mutagenesis and effects on enzyme activity were fluorometrically assayed. ## Methods Alanine substitutions were introduced by site-directed mutagenesis at residues L115, D129, G133, T134, Y150, G151, N152, S163 and I165 and recombinant proteins were purified

Who reads Structure-guided mutagenesis of active site residues in the dengue virus two-component protease NS2B-NS3?

Author

Wanisa Salaemae; Muhammad Junaid; Chanan Angsuthanasombat; Gerd Katzenmeier

Publisher

BioMed Central; Springer (Biomed Central Ltd.); BioMed Central Ltd.; [Basel Switzerland]: Karger; London: 2009-; Springer Science and Business Media LLC; S. Karger AG; Society for Mining, Metallurgy and Exploration Inc. (ISSN 1021-7770)

Published

2010

Language

EN

Field

Medicine (Life Sciences)