title: The baculovirus cysteine protease has a cathepsin B-like S2-subsite specificity.
abstract: Autographa californica nuclear polyhedrosis virus (AcNPV) encodes a functional cysteine protease of the papain family which is expressed after infection in Spodoptera fruglperda Sf9 cells. The protease displays an inhibition profile typical for cysteine proteases and is highly active against synthetic peptide substrates. The pH optimum of the bell-shaped pH-activity curve is between 5.0 and 5.5. The best substrate tested is Z-Arg-Arg-MCA which is specific for cathepsin B. The specificity constant (Kcat/Km) of AcNPV protease for this substrate is approximately two times higher than for human cathepsin B. In contrast to human cathepsins, AcNPV protease does not exhibit a discriminating specificity towards neutral hydrophobic residues in the P2 position. These substrates are hydrolysed at a ten-fold lower rate than the P2 arginine containing substrate. The pH activity profile against the Z-Arg-Arg-MCA substrate reveals a pK of 5.35 which can be assigned to a glutamate residue in the S2 subsite pocket. Like in cathepsin B, this residue facilitates the binding of positively charged P2 residues in the primary binding pocket. In this respect, the AcNPV protease resembles cathepsin B more than cathepsins L and S.
authors.affiliation: Khepri Pharmaceuticals, Inc., South San Francisco, CA 94080, USA.
authors.forename: D
authors.initials: D
authors.lastname: Brömme
authors.forename: K
authors.initials: K
authors.lastname: Okamoto
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002401:Cathepsin B / Q000737:chemistry*; D003546:Cysteine Endopeptidases / Q000737:chemistry* / Q000302:isolation & purification; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008969:Molecular Sequence Data; D017924:Nucleopolyhedroviruses / Q000201:enzymology* / Q000235:genetics; D018411:Spodoptera; D013379:Substrate Specificity
chemical_list: D015853:Cysteine Proteinase Inhibitors; D003546:Cysteine Endopeptidases; D002401:Cathepsin B