title: Human immunodeficiency virus type 1 protease inhibitors: evaluation of resistance engendered by amino acid substitutions in the enzyme's substrate binding site.
abstract: The human immunodeficiency virus type 1 (HIV-1) protease is a homodimeric aspartyl endopeptidase that is required for virus replication. A number of specific, active-site inhibitors for this enzyme have been described. Many of the inhibitors exhibit significant differences in activity against the HIV-1 and HIV type 2 (HIV-2) enzymes. An initial study was conducted to ascertain the HIV-1 protease's potential to lose sensitivity to several test inhibitors while retaining full enzymatic activity. The substrate binding sites of the HIV-1 and HIV-2 enzymes are almost fully conserved, except for four amino acid residues at positions 32, 47, 76, and 82. Accordingly, recombinant mutant type 1 proteases were constructed that contained the cognate type 2 residue at each of these four positions. The substitution at position 32 resulted in a significant adverse effect on inhibitor potency. However, this substitution also mediated a noted increase in the Km of the substrate. Individual substitutions at the remaining three positions, as well as a combination of all four substitutions, had very little effect on enzyme activity or inhibitor susceptibility. Hence, the four studied active site residues are insufficient to be responsible for differences in inhibitor sensitivity between the HIV-1 and HIV-2 proteases and are unlikely to contribute to the generation of inhibitor-resistant mutant HIV-1 protease.
authors.affiliation: Department of Virus and Cell Biology, Merck Research Laboratories, West Point, Pennsylvania 19486.
authors.forename: V V
authors.initials: VV
authors.lastname: Sardana
authors.forename: A J
authors.initials: AJ
authors.lastname: Schlabach
authors.forename: P
authors.initials: P
authors.lastname: Graham
authors.forename: B L
authors.initials: BL
authors.lastname: Bush
authors.forename: J H
authors.initials: JH
authors.lastname: Condra
authors.forename: J C
authors.initials: JC
authors.lastname: Culberson
authors.forename: L
authors.initials: L
authors.lastname: Gotlib
authors.forename: D J
authors.initials: DJ
authors.lastname: Graham
authors.forename: N E
authors.initials: NE
authors.lastname: Kohl
authors.forename: R L
authors.initials: RL
authors.lastname: LaFemina
mesh_terms: D000596:Amino Acids / Q000235:genetics*; D001578:Benzopyrans / Q000494:pharmacology; D001665:Binding Sites / Q000235:genetics; D003001:Cloning, Molecular; D003196:Computer Graphics; D004352:Drug Resistance, Microbial; D004926:Escherichia coli / Q000235:genetics; D016333:HIV Protease / Q000235:genetics* / Q000378:metabolism; D017320:HIV Protease Inhibitors / Q000737:chemistry / Q000494:pharmacology*; D007546:Isoquinolines / Q000494:pharmacology; D007700:Kinetics; D008958:Models, Molecular; D009025:Morpholines / Q000494:pharmacology; D010455:Peptides / Q000494:pharmacology; D011804:Quinolines / Q000494:pharmacology; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D019258:Saquinavir; D014633:Valine / Q000031:analogs & derivatives / Q000494:pharmacology
chemical_list: D000596:Amino Acids; D001578:Benzopyrans; D017320:HIV Protease Inhibitors; D007546:Isoquinolines; C086072:L 731723; D009025:Morpholines; D010455:Peptides; D011804:Quinolines; D011994:Recombinant Proteins; C072111:L 689502; D016333:HIV Protease; D014633:Valine; D019258:Saquinavir