title: Inhibiting rotavirus infection by membrane-impermeant thiol/disulfide exchange blockers and antibodies against protein disulfide isomerase.
abstract: OBJECTIVE
  Determining the effect of membrane-impermeant thiol/disulfide exchange inhibitors on rhesus rotavirus infectivity in MA104 cells and investigating protein disulfide isomerase (PDI) as a potential target for these inhibitors.
  
  
  METHODS
  Cells were treated with DTNB [5,5-dithio-bis-(2-nitrobenzoic acid)], bacitracin or anti-PDI antibodies and then infected with virus. Triple-layered particles (TLPs) were also pretreated with inhibitors before inoculation. The effects of these inhibitors on α-sarcin co-entry, virus binding to cells and PDI-TLP interaction were also examined. FACS analysis, cell-surface protein biotin-labeling, lipid-raft isolation and ELISA were performed to determine cell-surface PDI expression.
  
  
  RESULTS
  Infectivity became reduced by 50% when cells or TLPs were treated with 1 or 6 mM DTNB, respectively; infectivity became reduced by 50% by 20 mM bacitracin treatment of cells whereas TLPs were insensitive to bacitracin treatment; anti-PDI antibodies decreased viral infectivity by about 45%. The presence of DTNB (2.5 mM) or bacitracin (20 mM) was unable to prevent virus binding to cells and rotavirus-induced α-sarcin co-entry.
  
  
  CONCLUSIONS
  It was concluded that thiol/disulfide exchange was involved in rotavirus entry process and that cell-surface PDI was at least a potential target for DTNB and bacitracin-induced infectivity inhibition.
authors.affiliation: Chemistry Department, Science Faculty, Universidad Nacional de Colombia, Bogotá, Colombia.
authors.forename: Martha N
authors.initials: MN
authors.lastname: Calderon
authors.forename: Carlos A
authors.initials: CA
authors.lastname: Guerrero
authors.forename: Orlando
authors.initials: O
authors.lastname: Acosta
authors.forename: Susana
authors.initials: S
authors.lastname: Lopez
authors.forename: Carlos F
authors.initials: CF
authors.lastname: Arias
mesh_terms: D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D000906:Antibodies / Q000276:immunology; D001414:Bacitracin / Q000494:pharmacology; D002460:Cell Line; D004220:Disulfides / Q000378:metabolism; D004228:Dithionitrobenzoic Acid / Q000494:pharmacology; D004722:Endoribonucleases / Q000378:metabolism; D005656:Fungal Proteins / Q000378:metabolism; D008253:Macaca mulatta; D019704:Protein Disulfide-Isomerases / Q000037:antagonists & inhibitors* / Q000276:immunology / Q000378:metabolism; D012401:Rotavirus / Q000187:drug effects* / Q000502:physiology*; D012400:Rotavirus Infections / Q000188:drug therapy* / Q000378:metabolism; D013438:Sulfhydryl Compounds / Q000378:metabolism; D013439:Sulfhydryl Reagents / Q000494:pharmacology; D053585:Virus Attachment / Q000187:drug effects*; D053586:Virus Internalization / Q000187:drug effects*
chemical_list: D000900:Anti-Bacterial Agents; D000906:Antibodies; D004220:Disulfides; D005656:Fungal Proteins; D013438:Sulfhydryl Compounds; D013439:Sulfhydryl Reagents; D001414:Bacitracin; C014801:alpha-sarcin; D004228:Dithionitrobenzoic Acid; D004722:Endoribonucleases; D019704:Protein Disulfide-Isomerases