title: Identification of the galactose binding domain of the adeno-associated virus serotype 9 capsid.
abstract: Adeno-associated virus serotype 9 (AAV9) vectors show promise for gene therapy of a variety of diseases due to their ability to transduce multiple tissues, including heart, skeletal muscle, and the alveolar epithelium of the lung. In addition, AAV9 is unique compared to other AAV serotypes in that it is capable of surpassing the blood-brain barrier and transducing neurons in the brain and spinal cord. It has recently been shown that AAV9 uses galactose as a receptor to transduce many different cell types in vitro, as well as cells of the mouse airway in vivo. In this study, we sought to identify the specific amino acids of the AAV9 capsid necessary for binding to galactose. By site-directed mutagenesis and cell binding assays, plus computational ligand docking studies, we discovered five amino acids, including N470, D271, N272, Y446, and W503, which are required for galactose binding that form a pocket at the base of the protrusions around the icosahedral 3-fold axes of symmetry. The importance of these amino acids for tissue tropism was also confirmed by in vivo studies in the mouse lung. Identifying the interactions necessary for AAV9 binding to galactose may lead to advances in vector engineering.
authors.affiliation: Gene Therapy Program, Department of Pathology and Laboratory Medicine, Perelman School of Medicine at the University of Pennsylvania, Philadelphia, Pennsylvania, USA.
authors.forename: Christie L
authors.initials: CL
authors.lastname: Bell
authors.forename: Brittney L
authors.initials: BL
authors.lastname: Gurda
authors.forename: Kim
authors.initials: K
authors.lastname: Van Vliet
authors.forename: Mavis
authors.initials: M
authors.lastname: Agbandje-McKenna
authors.forename: James M
authors.initials: JM
authors.lastname: Wilson
mesh_terms: D019943:Amino Acid Substitution; D000818:Animals; D001665:Binding Sites; D016466:CHO Cells; D036022:Capsid Proteins / Q000235:genetics* / Q000378:metabolism*; D006224:Cricetinae; D000229:Dependovirus / Q000235:genetics* / Q000502:physiology; D005690:Galactose / Q000378:metabolism*; D008168:Lung / Q000821:virology; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D016297:Mutagenesis, Site-Directed; D050505:Mutant Proteins / Q000235:genetics / Q000378:metabolism; D053585:Virus Attachment
chemical_list: D036022:Capsid Proteins; D050505:Mutant Proteins; C018315:hexon capsid protein, Adenovirus; D005690:Galactose