Adenovirus endopeptidase and papain are inhibited by the same agents.
Adenoviruses encode a cysteine protease (AVP) which carries out highly specific cleavages on at least seven viral proteins and two cellular proteins. Virus infectivity is dependent on this function. The three-dimensional positions of the amino acids involved in catalysis display a striking similarity to those of papain, suggesting a similar catalytic mechanism. This similarity has prompted us to compare the effect of papain inhibitors on the two enzymes. AVP and papain activity was tested on a fluorescent peptide substrate as well as on metabolically labeled adenovirus (Ad2) precursor proteins. Hep2 cells infected with Ad2 were exposed to inhibitors and assayed for, (a) infectious virus, (b) in situ Ad2 protease activity, (c) physical particle production and their polypeptide composition. We found that in both substrate systems AVP was sensitive to the papain inhibitors benzamidoacetonitrile, acetamidoacetonitrile and N-methoxyphenylalanine glycylnitrile, and that the degree of sensitivity was influenced by the substrate. Unlike papain, AVP was relatively insensitive to E64. In ex vivo tests, Hep2 cells infected with Ad2 were exposed to inhibitors and assayed for, (a) infectious virus, (b) in situ Ad2 protease activity, (c) physical particle production and their polypeptide composition. A 4-fold reduction in virus titer was obtained when the inhibitors were added between 17 and 25 h after infection. Processing of precursor proteins was also inhibited yet the production of physical particles was only reduced 2-fold. These experiments show that papain inhibitors are also capable of inhibiting the adenovirus protease both in vitro and ex vivo, thus forging a possible link between structural similarity and functionality.
Sircar
S
S
Department of Microbiology, Faculty of Medicine, University of Sherbrooke, Que., Canada.
Ruzindana-Umunyana
A
A
Neugebauer
W
W
Weber
J M
JM
D000097:Acetonitriles / Q000494:pharmacology*; D000260:Adenoviruses, Human / Q000187:drug effects*; D000998:Antiviral Agents / Q000494:pharmacology*; D002384:Catalysis; D003546:Cysteine Endopeptidases / Q000378:metabolism*; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology*; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D006801:Humans; D009570:Nitriles / Q000494:pharmacology*; D010206:Papain / Q000037:antagonists & inhibitors*; D014407:Tumor Cells, Cultured
D000097:Acetonitriles; D000998:Antiviral Agents; D015853:Cysteine Proteinase Inhibitors; D004791:Enzyme Inhibitors; C117767:N-methoxyphenylalanine glycylnitrile; D009570:Nitriles; C117766:acetamidoacetonitrile; C117765:benzamidoacetonitrile; D003546:Cysteine Endopeptidases; D010206:Papain; C091827:adenain