In vivo and in vitro autoprocessing of human immunodeficiency virus protease expressed in Escherichia coli.
The viral-specific protease of human immunodeficiency virus (HIV) has been expressed as a lacZ-protease fusion protein. This fusion protein contains protease cleavage sites at the gag/protease and protease/reverse transcriptase junctions and undergoes autoprocessing in vivo when expressed in Escherichia coli. The purified lacZ-protease fusion protein precursors also exhibit autoproteolytic activity in vitro. One cleavage product of the autoprocessing reactions is a 10-kDa protein that cross-reacts with peptide antisera prepared against the putative protease sequence. Consistent with the notion that HIV protease is an acid protease, its autoproteolytic activity is inhibited in alkaline buffers and by pepstatin A. The in vivo and in vitro autocleavage assays for HIV protease together with the overproduction of the protease should facilitate design and testing of therapeutic agents that inhibit gag-pol polyprotein processing and HIV virion maturation.
Giam
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CZ
Department of Biochemistry, University of Nebraska Medical Center, College of Medicine, Omaha.
Boros
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D004262:DNA Restriction Enzymes; D004274:DNA, Recombinant / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics*; D005796:Genes; D005814:Genes, Viral; D006678:HIV / Q000201:enzymology / Q000235:genetics*; D008970:Molecular Weight; D010447:Peptide Hydrolases / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification; D011401:Promoter Regions, Genetic; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000302:isolation & purification
D004274:DNA, Recombinant; D011993:Recombinant Fusion Proteins; D004262:DNA Restriction Enzymes; D010447:Peptide Hydrolases