title: Generation and biological properties of a recombinant dodecahedron containing the short fiber protein of the human adenovirus 41.
abstract: OBJECTIVE
  In order to gain further insight into the function of the enteric adenovirus short fiber (SF), we have constructed a recombinant dodecahedron containing the SF protein of HAdV-41 and the HAdV-3 penton base.
  
  
  METHODS
  Recombinant baculoviruses expressing the HAdV-41 SF protein and HAdV-3 penton base were cloned and amplified in Sf9 insect cells. Recombinant dodecahedra were expressed by coinfection of High Five™ cells with both baculoviruses, 72 h post-infection. Cell lysate was centrifuged on sucrose density gradient and the purified recombinant dodecahedra were recovered.
  
  
  RESULTS
  Analysis by negative staining electron microscopy demonstrated that chimeric dodecahedra made of the HAdV-3 penton base and decorated with the HAdV-41 SF were successfully generated. Next, recombinant dodecahedra were digested with pepsin and analyzed by Western blot. A 'site-specific' proteolysis of the HAdV-41 SF was observed, while the HAdV-3 penton base core was completely digested.
  
  
  CONCLUSIONS
  These results show that, in vitro, the HAdV-41 SF likely undergoes proteolysis in the gastrointestinal tract, its natural environment, which may facilitate the recognition of receptors in intestinal cells. The results obtained in the present study may be the basis for the development of gene therapy vectors towards the intestinal epithelium, as well as orally administered vaccine vectors, but also for the HAdV-41 SF partner identification.
authors: Instituto de Ciências, Universidade de São Paulo, São Paulo, Brasil.
authors: Joselma
authors: J
authors: Siqueira-Silva
authors: Daphna
authors: D
authors: Fenel
authors: Evelyne
authors: E
authors: Gout
authors: Fernanda P
authors: FP
authors: Yeda
authors: Juliana C
authors: JC
authors: Marinheiro
authors: Karina M
authors: KM
authors: Barrella
authors: Misael L
authors: ML
authors: Silva
authors: Guy
authors: G
authors: Schoehn
authors: Charlotte M
authors: CM
authors: Harsi
authors: Pascal
authors: P
authors: Fender
mesh_terms: D000260:Adenoviruses, Human / Q000235:genetics*; D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D036022:Capsid Proteins / Q000235:genetics* / Q000378:metabolism / Q000648:ultrastructure*; D002460:Cell Line; D003001:Cloning, Molecular; D005822:Genetic Vectors; D007313:Insecta; D046911:Macromolecular Substances / Q000378:metabolism / Q000648:ultrastructure*; D008854:Microscopy, Electron; D010434:Pepsin A; D055503:Protein Multimerization; D059748:Proteolysis; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism / Q000648:ultrastructure; D022701:Virosomes / Q000235:genetics* / Q000378:metabolism / Q000648:ultrastructure*
chemical_list: D036022:Capsid Proteins; D046911:Macromolecular Substances; D011994:Recombinant Proteins; D022701:Virosomes; C018315:hexon capsid protein, Adenovirus; D010434:Pepsin A