title: Synthetic HIV-2 protease cleaves the GAG precursor of HIV-1 with the same specificity as HIV-1 protease.
abstract: A 99-amino acid protein having the deduced sequence of the protease from human immunodeficiency virus type 2 (HIV-2) was synthesized by the solid phase method and tested for specificity. The folded peptide catalyzes specific processing of a recombinant 43-kDa GAG precursor protein (F-16) of HIV-1. Although the protease of HIV-2 shares only 48% amino acid identity with that of HIV-1, the HIV-2 enzyme exhibits the same specificity toward the HIV-1 GAG precursor. Fragments of 34, 32, 24, 10, and 9 kDa were generated from F-16 GAG incubated with the protease. N-terminal amino acid sequence analysis of proteolytic fragments indicate that cleavage sites recognized by HIV-2 protease are identical to those of HIV-1 protease. The verified cleavage sites in F-16 GAG appear to be processed independently, as indicated by the formation of the intermediate fragments P32 and P34 in nearly equal ratios. The site nearest the amino terminus is quite conserved between the two viral GAG proteins (...VSQNY-PIVQN...in HIV-1,...KGGNY-PVQHV...in HIV-2). In contrast, the putative second site (...IPFAA-AQQKG...) of HIV-2 GAG shares minimal sequence identity with site 2 of HIV-1 GAG (...SATIM-MQRGN...). These sequence variations in the substrates suggest higher order structural features that may influence recognition by the proteases. Pepstatin A inhibits HIV-2 protease, whereas 1,10-phenanthroline and phenylmethylsulfonylfluoride do not; these results are in agreement with the finding that proteases of HIV and other retroviruses are aspartyl proteases.
authors.affiliation: Syntex Research, Palo Alto, California 94304.
authors.forename: J C
authors.initials: JC
authors.lastname: Wu
authors.forename: S F
authors.initials: SF
authors.lastname: Carr
authors.forename: K
authors.initials: K
authors.lastname: Jarnagin
authors.forename: S
authors.initials: S
authors.lastname: Kirsher
authors.forename: J
authors.initials: J
authors.lastname: Barnett
authors.forename: J
authors.initials: J
authors.lastname: Chow
authors.forename: H W
authors.initials: HW
authors.lastname: Chan
authors.forename: M S
authors.initials: MS
authors.lastname: Chen
authors.forename: D
authors.initials: D
authors.lastname: Medzihradszky
authors.forename: D
authors.initials: D
authors.lastname: Yamashiro
mesh_terms: D000595:Amino Acid Sequence; D015683:Gene Products, gag / Q000378:metabolism*; D015497:HIV-1 / Q000201:enzymology* / Q000378:metabolism; D008969:Molecular Sequence Data; D010447:Peptide Hydrolases / Q000138:chemical synthesis / Q000378:metabolism*; D011498:Protein Precursors / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism; D012689:Sequence Homology, Nucleic Acid; D013379:Substrate Specificity
chemical_list: D015683:Gene Products, gag; D011498:Protein Precursors; D011994:Recombinant Proteins; D010447:Peptide Hydrolases