Functional, biochemical and structural characterization of Hepatitis C virus non-structural protein 5A
Date of Issue2008
School of Biological Sciences
Non-structural protein 5A protein (NS5A) of Hepatitis C virus (HCY) plays an important role in the regulation of viral replication, interferon resistance, and apoptosis. The detailed mechanism and molecular characteristics ofNS5A remains unclear. In this study we first aimed to perform biophysical and biochemical characterization of the full length HCY NS5A. To this end, an efficient bacterial expression system was established to express NS5A and the system allowed us to purify the full length NS5A protein. The CD analysis of the full length NS5A suggested that this protein mainly consists of β-sheet at secondary structural level. In vitro GST pull-down assay proved that NS5A is functionally active and is capable of binding to its biological partners such as NS5B polymerase and PKR, a cellular interferon-inducible serine/threonine specific protein kinase. NS5A comprises three domains. Recently the structure of the domain 1 has been determined, revealing a structural scaffold with a novel zinc-binding motif and a disulfide bond.