Kinetics, inhibition and oligomerization of Epstein-Barr virus protease.
Epstein-Barr virus (EBV) is an omnipresent human virus causing infectious mononucleosis and EBV associated cancers. Its protease is a possible target for antiviral therapy. We studied its dimerization and enzyme kinetics with two enzyme assays based either on the release of paranitroaniline or 7-amino-4-methylcoumarin from labeled pentapeptide (Ac-KLVQA) substrates. The protease is in a monomer-dimer equilibrium where only dimers are active. In absence of citrate the K(d) is 20 microM and drops to 0.2 microM in presence of 0.5M citrate. Citrate increases additionally the activity of the catalytic sites. The inhibitory constants of different substrate derived peptides and alpha-keto-amide based inhibitors, which have at best a K(i) of 4 microM, have also been evaluated.
Buisson
Marlyse
M
Institut de Virologie Moléculaire et Structurale, FRE 2854 CNRS-UJF, BP181, 38042 Grenoble Cedex 9, France.
Rivail
Lucie
L
Hernandez
Jean-François
JF
Jamin
Marc
M
Martinez
Jean
J
Ruigrok
Rob W H
RW
Burmeister
Wim P
WP
D002863:Chromogenic Compounds / Q000494:pharmacology; D019343:Citric Acid / Q000494:pharmacology; D003124:Colorimetry; D019281:Dimerization; D010450:Endopeptidases / Q000737:chemistry* / Q000378:metabolism*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004854:Herpesvirus 4, Human / Q000201:enzymology*; D007700:Kinetics; D020836:Protein Structure, Quaternary / Q000187:drug effects; D014764:Viral Proteins / Q000037:antagonists & inhibitors* / Q000737:chemistry* / Q000378:metabolism
D002863:Chromogenic Compounds; D004791:Enzyme Inhibitors; D014764:Viral Proteins; D019343:Citric Acid; D010450:Endopeptidases