title: [Interaction of rotavirus with protein disulfide isomerase in vitro and cell system].
abstract: BACKGROUND
  Rotavirus entry process involves a multi-step mechanism, the first of which is when the outermost viral proteins interact with four different integrins and Hsc70. Recently, rotavirus infection reportedly has been decreased after blocking cell surface protein disulfide isomerase (PDI). This suggested that this protein interacts with rotavirus during the entry process.
  
  
  OBJECTIVE
  The aim was to establish the rotavirus-PDI interaction in an in vitro system using PDI isolated from bovine liver, and in a cell system consisting of MA104 cells and mouse small intestinal villi.
  
  
  METHODS
  Protein disulfide isomerase was isolated from a bovine liver homogenate using anti-PDI antibodies coupled to agarose through hydrazone bonds. Purity of purified protein was assessed by SDS-PAGE and Western blot. The purified PDI was used to study its in vitro interaction with the rotavirus particles. This interaction was compared with that taking place in MA104 cells and small intestinal villi isolated from sucking mice ICR.
  
  
  RESULTS
  The purified PDI showed an electrophoretic homogeneity and was able to bind rotavirus particles in vitro. Rotavirus-PDI interaction was detected by capture ELISA using purified protein and rotavirus strains RRV and wild-type ECwt. Interaction between rotavirus particles and cellular PDI was detected by ELISA using cell lysates after virus inoculation.
  
  
  CONCLUSIONS
  Rotavirus-PDI interaction was demonstrated in vitro as well as inMA104 cells and intestinal villi from suckling mice.
authors.affiliation: Facultad de Ciencias, Facultad de Ciencias, Universidad Nacional de Colombia, Bogotá, D.C, Colombia.
authors.forename: Martha N
authors.initials: MN
authors.lastname: Calderón
authors.forename: Carlos A
authors.initials: CA
authors.lastname: Guerrero
authors.forename: Yohana
authors.initials: Y
authors.lastname: Domínguez
authors.forename: Eliana
authors.initials: E
authors.lastname: Garzón
authors.forename: Sandra M
authors.initials: SM
authors.lastname: Barreto
authors.forename: Orlando
authors.initials: O
authors.lastname: Acosta
mesh_terms: D000818:Animals; D000833:Animals, Suckling; D000906:Antibodies / Q000378:metabolism; D002417:Cattle; D002460:Cell Line; D050883:HSC70 Heat-Shock Proteins / Q000378:metabolism; D016023:Integrins / Q000378:metabolism; D007413:Intestinal Mucosa / Q000201:enzymology / Q000821:virology; D007421:Intestine, Small / Q000166:cytology / Q000378:metabolism; D051379:Mice; D019704:Protein Disulfide-Isomerases / Q000378:metabolism*; D012401:Rotavirus / Q000378:metabolism*; D012400:Rotavirus Infections / Q000821:virology; D014764:Viral Proteins / Q000378:metabolism
chemical_list: D000906:Antibodies; D050883:HSC70 Heat-Shock Proteins; D016023:Integrins; D014764:Viral Proteins; D019704:Protein Disulfide-Isomerases