title: Analysis of the interaction between adeno-associated virus and heparan sulfate using atomic force microscopy.
abstract: Adeno-associated virus (AAV) has been widely used as a viral vector to deliver genes to animal and human tissues in gene therapy studies. Both AAV-2 and AAV-3 use cell surface heparan sulfate (HS), a highly sulfated polysaccharide, as a receptor to establish infections. In this study, we used atomic force microscopy (AFM) to investigate the interaction of HS and AAV. A silicon chip functionalized with HS was used as a substrate for binding AAV for AFM analysis. To validate our approach, we found that the binding of AAV-2 to the HS surface was effectively competed by soluble HS, suggesting that the binding of AAV-2 to the functionalized surface was specific. In addition, we examined the binding of various AAV serotypes, including AAV-1, AAV-2, AAV-3, and AAV-5, to the HS surface. As expected, only AAV-2 and AAV-3 bound, whereas AAV-1 and AAV-5 did not. This observation was consistent with the previous conclusion that AAV-1 and AAV-5 do not use HS as a receptor for infection. In conclusion, we developed a novel approach to investigate the interaction of AAV virus with its polysaccharide-based receptor at the level of a single viral particle. Given that HSs serve as receptor for numerous viruses, this approach has the potential to become a generalized method for studying interactions between the viral particle and HS, as well as other virus-cell interactions, and potentially serve as a platform for screening antiviral therapies.
authors.affiliation: Curriculum in Applied and Materials Sciences, Program in Cellular and Molecular Biophysics, CB# 3287, University of North Carolina, Chapel Hill, NC 27599, USA.
authors.forename: Atsuko
authors.initials: A
authors.lastname: Negishi
authors.forename: Jinghua
authors.initials: J
authors.lastname: Chen
authors.forename: Douglas M
authors.initials: DM
authors.lastname: McCarty
authors.forename: R Jude
authors.initials: RJ
authors.lastname: Samulski
authors.forename: Jian
authors.initials: J
authors.lastname: Liu
authors.forename: Richard
authors.initials: R
authors.lastname: Superfine
mesh_terms: D002460:Cell Line; D002462:Cell Membrane / Q000821:virology; D000229:Dependovirus / Q000737:chemistry* / Q000648:ultrastructure*; D006497:Heparan Sulfate / Q000737:chemistry* / Q000648:ultrastructure*; D006801:Humans; D018625:Microscopy, Atomic Force; D012995:Solubility; D013501:Surface-Active Agents
chemical_list: D006497:Heparan Sulfate; D013501:Surface-Active Agents