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
| Autor: | Sudir Sumarheni, Pierre Boulanger, Pascal Fender, Jean-Luc Coll, Saw See Hong, Guy Schoehn, Véronique Josserand |
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| Přispěvatelé: | Department of Chemistry and Nano Science, EWHA Womans University (EWHA), Université Joseph Fourier - Grenoble 1 (UJF), Radiopharmaceutiques biocliniques (LRB), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019]), Institut d'oncologie/développement Albert Bonniot de Grenoble (INSERM U823), Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM), AMMI Lab, University of Alberta, Institut de biologie structurale (IBS - UMR 5075 ), Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS), 'Bio-Health Computing' European training program, FRISBI [ANR-10-INSB-05-02], GRAL [ANR-10-LABX-49-01], Rhone-Alpes Region, FRM, CNRS, University of Grenoble, GIS-IBISA, French Foundation for Cystic Fibrosis (Vaincre la Mucoviscidose) [VLM-RF2013-0500796], French Institute of Health and Medical Research (Institut National de la Sante et de la Recherche Medicale, INSERM), Contrat d'Interface INSERM-Hospices Civils de Lyon [CIF-20082013], Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA) |
| Rok vydání: | 2014 |
| Předmět: |
Models
Molecular Hexon binding Protein Conformation [SDV]Life Sciences [q-bio] viruses Gene Expression Peptide Plasma protein binding chemistry.chemical_compound Transduction (genetics) Mice 0302 clinical medicine Protein structure Genes Reporter Neoplasms Research Articles chemistry.chemical_classification 0303 health sciences Factor X virus diseases 030220 oncology & carcinogenesis Molecular Medicine Female hormones hormone substitutes and hormone antagonists Protein Binding Genetic Vectors Biology Binding Competitive Cell Line 03 medical and health sciences Genetics Animals Humans Protein Interaction Domains and Motifs Molecular Biology 030304 developmental biology Gla domain Adenoviruses Human Capsomere Virion Surface Plasmon Resonance Molecular biology eye diseases Disease Models Animal Viral Tropism chemistry Luminescent Measurements Capsid Proteins Peptides |
| Zdroj: | Human Gene Therapy Human Gene Therapy, Mary Ann Liebert, 2014, 25 (4), pp.339-49. ⟨10.1089/hum.2013.222⟩ Human Gene Therapy, 2014, 25 (4), pp.339-49. ⟨10.1089/hum.2013.222⟩ |
| ISSN: | 1557-7422 1043-0342 |
| DOI: | 10.1089/hum.2013.222⟩ |
| Popis: | International audience; 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. |
| Databáze: | OpenAIRE |
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