title: Human full-length coagulation factor X and a GLA domain-derived 40-mer polypeptide bind to different regions of the adenovirus serotype 5 hexon capsomer.
abstract: The interaction of human adenovirus (HAdV)-C5 and many other adenoviruses with blood coagulation factors (e.g., human factor X, FX) involves the binding of their GLA domain to the hexon capsomers, resulting in high levels of hepatotropism and potential hepatotoxicity. In this study, we tested the possibility of preventing these undesirable effects by using a GLA-mimicking peptide as a competitor. An FX GLA domain-derived, 40-mer polypeptide carrying 12 carboxyglutamate residues was synthesized (GLA(mim)). Surface plasmon resistance (SPR) analysis showed that GLA(mim) reacted with free and capsid-embedded hexon with a nanomolar affinity. Unexpectedly, GLA(mim) failed to compete with FX for hexon binding, and instead significantly increased the formation of FX-hexon or FX-adenovirion complexes. This observation was confirmed by in vitro cell transduction experiments using HAdV-C5-Luciferase vector (HAdV5-Luc), as preincubation of HAdV5-Luc with GLA(mim) before FX addition resulted in a higher transgene expression compared with FX alone. HAdV-C5 virions complexed with GLA(mim) were analyzed by cryoelectron microscopy. Image reconstruction demonstrated the bona fide hexon-GLA(mim) interaction, as for the full-length FX, although with considerable differences in stoichiometry and relative location on the hexon capsomer. Three extra densities were found at the periphery of each hexon, whereas one single FX molecule occupied the central cavity of the hexon trimeric capsomer. A refined analysis indicated that each extra density is found at the expected location of one highly variable loop 1 of the hexon, involved in scavenger receptor recognition. HAdV5-Luc complexed with a bifunctional GLA(mim)RGD peptide showed a lesser hepatotropism, compared with control HAdV5-Luc alone, and efficiently targeted αβ-integrin-overexpressing tumor cells in an in vivo mouse tumor model. Collectively, our findings open new perspectives in the design of adenoviral vectors for biotherapy.
authors: 1 Centre National Recherche Scientifique, Université Joseph Fourier , European Molecular Biology Laboratory (Unit of Virus Host Cell Interactions; Unité Mixte Internationale-3265), 38042 Grenoble, France .
authors: Sudir
authors: S
authors: Sumarheni
authors: Saw See
authors: SS
authors: Hong
authors: Véronique
authors: V
authors: Josserand
authors: Jean-Luc
authors: JL
authors: Coll
authors: Pierre
authors: P
authors: Boulanger
authors: Guy
authors: G
authors: Schoehn
authors: Pascal
authors: P
authors: Fender
mesh_terms: D000260:Adenoviruses, Human / Q000235:genetics* / Q000378:metabolism* / Q000648:ultrastructure; D000818:Animals; D001667:Binding, Competitive; D036022:Capsid Proteins / Q000737:chemistry / Q000378:metabolism*; D002460:Cell Line; D004195:Disease Models, Animal; D005170:Factor X / Q000737:chemistry / Q000235:genetics*; D005260:Female; D015870:Gene Expression; D017930:Genes, Reporter; D005822:Genetic Vectors / Q000235:genetics*; D006801:Humans; D008163:Luminescent Measurements; D051379:Mice; D008958:Models, Molecular; D009369:Neoplasms / Q000378:metabolism / Q000473:pathology; D010455:Peptides / Q000737:chemistry / Q000378:metabolism*; D011485:Protein Binding; D011487:Protein Conformation; D054730:Protein Interaction Domains and Motifs; D020349:Surface Plasmon Resonance; D056189:Viral Tropism; D014771:Virion / Q000648:ultrastructure
chemical_list: D036022:Capsid Proteins; D010455:Peptides; C018315:hexon capsid protein, Adenovirus; D005170:Factor X