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Blood, 30 July 2009, Vol. 114, No. 5, pp. 965-971. Prepublished online as a Blood First Edition Paper on May 8, 2009; DOI 10.1182/blood-2009-03-208835. This Article Full Text Full Text (PDF) Supplemental Table and Figures All Versions of this Article: blood-2009-03-208835v1 114/5/965    most recent Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Rights and Permissions Citing Articles Citing Articles via HighWire Citing Articles via CrossRef Google Scholar Articles by Alba, R. Articles by Baker, A. H. PubMed PubMed Citation Articles by Alba, R. Articles by Baker, A. H. Related Collections Thrombosis and Hemostasis Gene Therapy Related Article in Blood Online Social Bookmarking                   What's this?  Previous Article  |  Table of Contents  |  Next Article  GENE THERAPY Identification of coagulation factor (F)X binding sites on the adenovirus serotype 5 hexon: effect of mutagenesis on FX interactions and gene transfer Raul Alba1, Angela C. Bradshaw1, Alan L. Parker1, David Bhella2, Simon N. Waddington3, Stuart A. Nicklin1, Nico van Rooijen4, Jerome Custers5, Jaap Goudsmit5, Dan H. Barouch6, John H. McVey7, and Andrew H. Baker1 1 British Heart Foundation Glasgow Cardiovascular Research Centre, and 2 Medical Research Council (MRC) Virology Unit, University of Glasgow, Glasgow, United Kingdom; 3 Department of Haematology, Haemophilia Centre and Haemostasis Unit, Royal Free and University College Medical School, London, United Kingdom; 4 Department of Molecular Cell Biology, Vrije Universiteit Medical Center (VUMC), Amsterdam, The Netherlands; 5 Crucell Holland BV, Leiden, The Netherlands; 6 Beth Israel Deaconess Medical Center, Boston, MA; and 7 Thrombosis Research Institute, London, United Kingdom Recent studies have demonstrated the importance of coagulation factor X (FX) in adenovirus (Ad) serotype 5–mediated liver transduction in vivo. FX binds to the adenovirus hexon hypervariable regions (HVRs). Here, we perform a systematic analysis of FX binding to Ad5 HVRs 5 and 7, identifying domains and amino acids critical for this interaction. We constructed a model of the Ad5-FX interaction using crystallographic and cryo-electron microscopic data to identify contact points. Exchanging Ad5 HVR5 or HVR7 from Ad5 to Ad26 (which does not bind FX) diminished FX binding as analyzed by surface plasmon resonance, gene delivery in vitro, and liver transduction in vivo. Exchanging Ad5-HVR5 for Ad26-HVR5 produced deficient virus maturation. Importantly, defined mutagenesis of just 2 amino acids in Ad5-HVR5 circumvented this and was sufficient to block liver gene transfer. In addition, mutation of 4 amino acids in Ad5-HVR7 or a single mutation at position 451 also blocked FX-mediated effects in vitro and in vivo. We therefore define the regions and amino acids on the Ad5 hexon that bind with high affinity to FX thereby better defining adenovirus infectivity pathways. These vectors may be useful for gene therapy applications where evasion of liver transduction is a prerequisite. CiteULike    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this? Related Article in Blood Online: Mutagenesis of hexon "FX" hepatic tropism Eric J. Kremer Blood 2009 114: 929-930. [Abstract] [Full Text] [PDF] This article has been cited by other articles: E. J. Kremer Mutagenesis of hexon "FX" hepatic tropism Blood, July 30, 2009; 114(5): 929 - 930. [Abstract] [Full Text] [PDF]

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