title: Activity of purified biosynthetic proteinase of human immunodeficiency virus on natural substrates and synthetic peptides.
abstract: Retroviral capsid proteins and replication enzymes are synthesized as polyproteins that are proteolytically processed to the mature products by a virus-encoded proteinase. We have purified the proteinase of human immunodeficiency virus (HIV), expressed in Escherichia coli, to approximately 90% purity. The purified enzyme at a concentration of approximately 20 nM gave rapid, efficient, and specific cleavage of an in vitro synthesized gag precursor protein. Purified HIV proteinase also induced specific cleavage of five decapeptide substrates whose amino acid sequences corresponded to cleavage sites in the HIV polyprotein but not of a peptide corresponding to a cleavage site in another retrovirus. Competition experiments with different peptides allowed a ranking of cleavage sites. Inhibition studies indicated that the HIV proteinase was inhibited by pepstatin A with an IC50 of 0.7 microM.
authors.affiliation: Department of Microbiology, State University of New York, Stony Brook 11794.
authors.forename: H G
authors.initials: HG
authors.lastname: Kräusslich
authors.forename: R H
authors.initials: RH
authors.lastname: Ingraham
authors.forename: M T
authors.initials: MT
authors.lastname: Skoog
authors.forename: E
authors.initials: E
authors.lastname: Wimmer
authors.forename: P V
authors.initials: PV
authors.lastname: Pallai
authors.forename: C A
authors.initials: CA
authors.lastname: Carter
mesh_terms: D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D006678:HIV / Q000201:enzymology* / Q000235:genetics; D007700:Kinetics; D010447:Peptide Hydrolases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D010957:Plasmids; D014176:Protein Biosynthesis; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism*; D013379:Substrate Specificity; D014158:Transcription, Genetic; D014764:Viral Proteins
chemical_list: D011994:Recombinant Proteins; D014764:Viral Proteins; D010447:Peptide Hydrolases