Type of Document	Dissertation
Author	O'Donnell, Jason Kelly
URN	etd-07262009-152850
Title	Adeno-Associated Virus-2 and its Primary Cellular Receptor-Cryo-EM Structure of a Heparin Complex
Degree	Doctor of Philosophy
Department	Chemistry and Biochemistry, Department of
Advisory Committee	Advisor Name Title Hong Li Committee Co-Chair Michael Chapman Committee Co-Chair Sanford Safron Committee Member Kenneth Taylor Outside Committee Member
Keywords	Virus Cryo-EM Single Particle Analysis Electron Microscopy Receptor
Date of Defense	2009-05-07
Availability	unrestricted
Abstract Adeno-associated virus serotype 2 (AAV-2) is a leading candidate vector for gene therapy. Cell entry starts with attachment to a primary receptor, Heparan Sulfate Proteoglycan (HSPG) before binding to a co-receptor. Here, cryo-electron microscopy provides direct visualization of the virus–HSPG interactions. Single particle analysis was performed on AAV-2 complexed with a 17 kDa heparin fragment at 8.3 Å resolution. Heparin density covers the shoulder of spikes surrounding viral 3-fold symmetry axes. Previously implicated, positively charged residues R448/585, R451/588 and R350/487 from another subunit cluster at the center of the heparin footprint. The footprint is much more extensive than apparent through mutagenesis, including R347/484, K395/532 and K390/527 that are more conserved, but whose roles have been controversial. It also includes much of a region proposed as a co-receptor site, because prior studies had not revealed heparin interactions. Heparin density bridges over the viral 3-fold axes, indicating multi-valent attachment to symmetry-related binding sites.
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