title: Blocking human enterovirus 71 replication by targeting viral 2A protease.
abstract: OBJECTIVE
  Human enterovirus 71 (EV-71), a member of the Enterovirus genus, constitutes a major public health issue in the Asia-Pacific region, where it is associated with several severe neurological complications. There is currently no effective vaccine or antiviral against EV-71. The aim of this study was to determine whether the six amino acid peptide LVLQTM, which was previously shown to inhibit human rhinovirus (HRV) 2A protease (2A(pro)) activity in vitro and HRV replication in vivo in mice, could be of more general use against enteroviruses and more particularly against EV-71.
  
  
  METHODS
  To investigate whether the LVLQTM peptide was a pseudosubstrate of EV-71 2A(pro), a recombinant luciferase containing the LVLQTM sequence was designed so that recognition of this sequence by 2A(pro) led to luciferase activation. Direct interaction between EV-71 2A(pro) and the LVLQTM peptide was further confirmed by isothermal titration calorimetry. We then tested the effects of the peptide on EV-71 2A(pro) cleavage activity and EV-71 replication in HeLa cells.
  
  
  RESULTS
  We showed that the LVLQTM peptide behaved as an effective substrate analogue of EV-71 2A(pro), which binds into the active site of the protease with a dissociation rate constant of 9.6 μM. Moreover, LVLQTM significantly inhibited eIF4G cleavage activity of 2A(pro) as well as EV-71 replication in HeLa cells.
  
  
  CONCLUSIONS
  This study demonstrates that the LVLQTM peptide that has previously been shown to inhibit HRV replication is also an effective inhibitor of EV-71 2A(pro) and therefore of EV-71 replication, opening new doors in the development of new antivirals against EV-71.
authors.affiliation: VirPath, Virologie et Pathologie Humaine, EA4610, Université Lyon 1, Université de Lyon, Faculté de Médecine Lyon-Est, secteur Laennec,  Rue Guillaume Paradin, F-69372 Lyon cedex 08,  France.
authors.forename: Nisrine
authors.initials: N
authors.lastname: Falah
authors.forename: Roland
authors.initials: R
authors.lastname: Montserret
authors.forename: Virginie
authors.initials: V
authors.lastname: Lelogeais
authors.forename: Isabelle
authors.initials: I
authors.lastname: Schuffenecker
authors.forename: Bruno
authors.initials: B
authors.lastname: Lina
authors.forename: Jean-Claude
authors.initials: JC
authors.lastname: Cortay
authors.forename: Sébastien
authors.initials: S
authors.lastname: Violot
mesh_terms: D000998:Antiviral Agents / Q000494:pharmacology*; D003546:Cysteine Endopeptidases / Q000378:metabolism*; D029821:Enterovirus A, Human / Q000187:drug effects / Q000201:enzymology* / Q000502:physiology*; D006367:HeLa Cells; D006801:Humans; D009842:Oligopeptides / Q000494:pharmacology; D011480:Protease Inhibitors / Q000494:pharmacology*; D011485:Protein Binding; D014764:Viral Proteins / Q000378:metabolism*; D014779:Virus Replication / Q000187:drug effects*
chemical_list: D000998:Antiviral Agents; D009842:Oligopeptides; D011480:Protease Inhibitors; D014764:Viral Proteins; D003546:Cysteine Endopeptidases