title: Potent inhibition of drug-resistant HIV protease variants by monoclonal antibodies.
abstract: The monoclonal antibodies 1696 and F11.2.32 strongly inhibit the activity of wild-type HIV-1 protease (PR) by binding to epitopes at the enzyme N-terminus (residues 1-6) and flap residues 36-46, respectively. Here we demonstrate that these antibodies are also potent inhibitors of PR variants resistant to active-site inhibitors used as anti-AIDS drugs. Our in vitro experiments revealed that the inhibitory potency of single-chain fragments (scFv) of these antibodies is not significantly affected by the presence of mutations in PR; inhibition constants for drug-resistant protease variants are 5-11 nM and 13-169 nM for scFv1696 and for scFvF11.2.32, respectively. Tethered dimer of HIV-1 PR variant proved to be a model protease variant resistant to dissociative inhibition by 1696, and, strikingly, it also displayed resistance to inhibition by F11.2.32 suggesting that dimer dissociation also plays a role in the inhibitory action of F11.2.32.
authors.affiliation: Institute of Molecular Genetics, Academy of Sciences of the Czech Republic, Flemingovo nam. 2, 166 37 Prague 6, Czech Republic.
authors.forename: Vanda
authors.initials: V
authors.lastname: Bartonová
authors.forename: Vlastimil
authors.initials: V
authors.lastname: Král
authors.forename: Irena
authors.initials: I
authors.lastname: Sieglová
authors.forename: Jirí
authors.initials: J
authors.lastname: Brynda
authors.forename: Milan
authors.initials: M
authors.lastname: Fábry
authors.forename: Magdalena
authors.initials: M
authors.lastname: Horejsí
authors.forename: Milan
authors.initials: M
authors.lastname: Kozísek
authors.forename: Klára Grantz
authors.initials: KG
authors.lastname: Sasková
authors.forename: Jan
authors.initials: J
authors.lastname: Konvalinka
authors.forename: Juraj
authors.initials: J
authors.lastname: Sedlácek
authors.forename: Pavlína
authors.initials: P
authors.lastname: Rezácová
mesh_terms: D000911:Antibodies, Monoclonal / Q000276:immunology / Q000494:pharmacology*; D023241:Antiretroviral Therapy, Highly Active; D019281:Dimerization; D024882:Drug Resistance, Viral / Q000235:genetics*; D014644:Genetic Variation; D015658:HIV Infections / Q000188:drug therapy / Q000821:virology; D016333:HIV Protease / Q000187:drug effects* / Q000235:genetics / Q000276:immunology; D017320:HIV Protease Inhibitors / Q000494:pharmacology; D015497:HIV-1 / Q000187:drug effects / Q000201:enzymology* / Q000235:genetics; D006801:Humans; D007128:Immunoglobulin Fragments / Q000276:immunology / Q000494:pharmacology*; D008958:Models, Molecular; D009154:Mutation; D011994:Recombinant Proteins / Q000276:immunology / Q000494:pharmacology*
chemical_list: D000911:Antibodies, Monoclonal; D017320:HIV Protease Inhibitors; D007128:Immunoglobulin Fragments; D011994:Recombinant Proteins; C055966:immunoglobulin Fv; D016333:HIV Protease