Characterization of the murine leukemia virus protease and its comparison with the human immunodeficiency virus type 1 protease.
The protease (PR) of Murine leukemia virus (MLV) was expressed in Escherichia coli, purified to homogeneity and characterized by using various assay methods, including HPLC-based, photometric and fluorometric activity measurements. The specificity of the bacterially expressed PR was similar to that of virion-extracted PR. Compared with human immunodeficiency virus type 1 (HIV-1) PR, the pH optimum of the MLV enzyme was higher. The specificity of the MLV PR was further compared with that of HIV-1 PR by using various oligopeptides representing naturally occurring cleavage sites in MLV and HIV-1, as well as by using bacterially expressed proteins having part of the MLV Gag. Inhibitors designed against HIV-1 PR were also active on MLV PR, although all of the tested ones were substantially less potent on this enzyme than on HIV-1 PR. Nevertheless, amprenavir, the most potent inhibitor against MLV PR, was also able to block Gag processing in MLV-infected cells. These results indicate that, in spite of the similar function in the life cycle of virus infection, the two PRs are only distantly related in their specificity.
Fehér
Anita
A
Department of Biochemistry and Molecular Biology, Research Center for Molecular Medicine, Medical and Health Science Center, University of Debrecen, Hungary.
Boross
Péter
P
Department of Biochemistry and Molecular Biology, Research Center for Molecular Medicine, Medical and Health Science Center, University of Debrecen, Hungary.
Sperka
Tamás
T
Department of Biochemistry and Molecular Biology, Research Center for Molecular Medicine, Medical and Health Science Center, University of Debrecen, Hungary.
Miklóssy
Gabriella
G
Department of Biochemistry and Molecular Biology, Research Center for Molecular Medicine, Medical and Health Science Center, University of Debrecen, Hungary.
Kádas
János
J
Department of Biochemistry and Molecular Biology, Research Center for Molecular Medicine, Medical and Health Science Center, University of Debrecen, Hungary.
Bagossi
Péter
P
Department of Biochemistry and Molecular Biology, Research Center for Molecular Medicine, Medical and Health Science Center, University of Debrecen, Hungary.
Oroszlan
Stephen
S
HIV Drug Resistant Program, National Cancer Institute at Frederick, MD, USA.
Weber
Irene T
IT
Department of Biology, Georgia State University, Atlanta, GA, USA.
Tözsér
József
J
Department of Biochemistry and Molecular Biology, Research Center for Molecular Medicine, Medical and Health Science Center, University of Debrecen, Hungary.
D000595:Amino Acid Sequence; D000818:Animals; D002219:Carbamates / Q000494:pharmacology; D004926:Escherichia coli / Q000378:metabolism; D005663:Furans; D015683:Gene Products, gag / Q000235:genetics / Q000378:metabolism; D016333:HIV Protease / Q000378:metabolism; D017320:HIV Protease Inhibitors / Q000494:pharmacology; D006801:Humans; D006863:Hydrogen-Ion Concentration; D009052:Leukemia Virus, Murine / Q000201:enzymology* / Q000276:immunology; D051379:Mice; D008969:Molecular Sequence Data; D041681:NIH 3T3 Cells; D009842:Oligopeptides / Q000378:metabolism; D010447:Peptide Hydrolases / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000378:metabolism; D013045:Species Specificity; D013449:Sulfonamides / Q000494:pharmacology
D002219:Carbamates; D005663:Furans; D015683:Gene Products, gag; D017320:HIV Protease Inhibitors; D009842:Oligopeptides; D011994:Recombinant Proteins; D013449:Sulfonamides; C095108:amprenavir; D010447:Peptide Hydrolases; D016333:HIV Protease