Kinetic characterization of human immunodeficiency virus type 1 protease: determination of inhibitor rate constants during dynamic monomer-dimer interconversion.
A numerical method was applied to a system of differential rate equations describing the monomer-dimer-inhibitor (M-D-I) interaction involving human immunodeficiency virus type 1 protease and a peptidomimetic, competitive inhibitor. Two pairs of progress curves were obtained, one involving the M-D interaction and the other the M-D-I interaction. Each pair of reactions was designed to begin with extreme conditions and end at the identical equilibrium position. The results were compared with analytical (exact mathematical) methods reported previously. Good agreement between the two methods was observed at high- and low-salt conditions for the rates of monomer association and dimer dissociation. Not surprisingly, however, the major difference was observed in the analyses involving the M-D-I interaction, since analytical methods cannot account for dimer dissociation in the presence of inhibitor. While the estimates for the inhibitor off rate were comparable for high-salt conditions (where dimer dissociation is minimized), the analytical method underestimated this parameter for low-salt conditions by an order of magnitude, the consequence of mistaking inactive M for inactive DI.
Morelock
M M
MM
Section of Biophysics, Department of Inflammatory Diseases, Boehringer Ingelheim Pharmaceuticals, Inc., Research and Development Center, Ridgefield, Connecticut 06877-0368, USA.
Graham
E T
ET
Erdman
D
D
Pargellis
C A
CA
D001667:Binding, Competitive; D004926:Escherichia coli; D016333:HIV Protease / Q000737:chemistry* / Q000378:metabolism*; D017320:HIV Protease Inhibitors / Q000494:pharmacology*; D015497:HIV-1 / Q000201:enzymology*; D006801:Humans; D007700:Kinetics; D046911:Macromolecular Substances; D008433:Mathematics; D008962:Models, Theoretical; D011994:Recombinant Proteins / Q000378:metabolism
D017320:HIV Protease Inhibitors; D046911:Macromolecular Substances; D011994:Recombinant Proteins; D016333:HIV Protease