CpAMs induce assembly of HBV capsids with altered electrophoresis mobility: Implications for mechanism of inhibiting pgRNA packaging.

摘要: Native agarose gel electrophoresis-based particle assay has been commonly used for examination of hepatitis B virus (HBV) capsid assembly and pregenomic RNA encapsidation in HBV replicating cells. Interestingly, treatment cells with several chemotypes core protein allosteric modulators (CpAMs) induced the both empty DNA-containing capsids faster electrophoresis mobility. In an effort to determine physical basis CpAM-induced mobility shift, we found that surface charge, but not size, is primary determinant Specifically, through alanine scanning mutagenesis analysis twenty-seven charged amino acids domain hinge region, showed except K7 E8, substitution glutamine acid (E) or aspartic (D) on reduced their mobility, lysine (K) arginine (R) increased variable degrees. However, are exposed did apparently alter Hence, shift may reflect global alteration structure changes exposure and/or ionization side chains protein. Our findings imply CpAM inhibition pgRNA possibly due structurally altered nucleocapsids. Practically, a diagnostic marker compounds target predicts sensitivity strains specific CpAMs.