Abstract
We performed a mutational analysis of the NS3 helicase of dengue virus to test insights gleaned from its crystal structure and identified four residues in the full-length protein that severely impaired either its RTPase and ATPase (Arg-457-458, Arg-460, Arg-463) or helicase (Ile-365, Arg-376) activity. Alanine substitution of Lys-396, which is located at the surface of domain II, drastically reduced all three enzymatic activities. Our study points to a pocket at the surface of domain II that may be suitable for the design of allosteric inhibitors.
MeSH terms
-
Allosteric Site / genetics
-
Amino Acid Substitution*
-
Drug Design
-
Enzyme Inhibitors / chemistry
-
Point Mutation*
-
Protein Structure, Tertiary / genetics
-
RNA Helicases / antagonists & inhibitors
-
RNA Helicases / chemistry
-
RNA Helicases / genetics
-
Serine Endopeptidases / chemistry
-
Serine Endopeptidases / genetics
-
Structure-Activity Relationship
-
Viral Nonstructural Proteins / antagonists & inhibitors
-
Viral Nonstructural Proteins / chemistry*
-
Viral Nonstructural Proteins / genetics
Substances
-
Enzyme Inhibitors
-
NS3 protein, flavivirus
-
Viral Nonstructural Proteins
-
Serine Endopeptidases
-
RNA Helicases