Adeno-associated virus-2 and its primary cellular receptor-Cryo-EM structure of a heparin complex

Jason O'Donnell; Kenneth A. Taylor; Michael S. Chapman

doi:10.1016/j.virol.2008.11.037

Adeno-associated virus-2 and its primary cellular receptor-Cryo-EM structure of a heparin complex

Jason O'Donnell, Kenneth A. Taylor, Michael S. Chapman

Research output: Contribution to journal › Article › peer-review

103 Scopus citations

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 R_448/585, R_451/588 and R_350/487 from another subunit cluster at the center of the heparin footprint. The footprint is much more extensive than apparent through mutagenesis, including R_347/484, K_395/532 and K_390/527 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.

Original language	English (US)
Pages (from-to)	434-443
Number of pages	10
Journal	Virology
Volume	385
Issue number	2
DOIs	https://doi.org/10.1016/j.virol.2008.11.037
State	Published - Mar 15 2009
Externally published	Yes

Keywords

Electron microscopy
Gene therapy
Heparan sulfate
Heparin
Parvovirus

ASJC Scopus subject areas

Virology

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10.1016/j.virol.2008.11.037

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title = "Adeno-associated virus-2 and its primary cellular receptor-Cryo-EM structure of a heparin complex",

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 {\AA} 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.",

keywords = "Electron microscopy, Gene therapy, Heparan sulfate, Heparin, Parvovirus",

author = "Jason O'Donnell and Taylor, {Kenneth A.} and Chapman, {Michael S.}",

note = "Funding Information: We wish to thank Joan Hare at the Protein expression facility for virus propagation and purification and Kim Riddle and Tom Fellers at the Biological Science Imaging resource for assistance in data collection. We gratefully acknowledge the computer resources and expertise provided by the High Performance Computing center at Florida State University. Molecular graphics images were produced using the UCSF Chimera package from the Resource for Biocomputing, Visualization, and Informatics at the University of California, San Francisco (supported by NIH P41 RR-01081). This work was supported by an American Heart Association pre-doctoral fellowship (J.O'D) and the National Institutes of Health (R01 GM 66875, M.S.C.).",

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doi = "10.1016/j.virol.2008.11.037",

language = "English (US)",

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T1 - Adeno-associated virus-2 and its primary cellular receptor-Cryo-EM structure of a heparin complex

AU - O'Donnell, Jason

AU - Taylor, Kenneth A.

AU - Chapman, Michael S.

N1 - Funding Information: We wish to thank Joan Hare at the Protein expression facility for virus propagation and purification and Kim Riddle and Tom Fellers at the Biological Science Imaging resource for assistance in data collection. We gratefully acknowledge the computer resources and expertise provided by the High Performance Computing center at Florida State University. Molecular graphics images were produced using the UCSF Chimera package from the Resource for Biocomputing, Visualization, and Informatics at the University of California, San Francisco (supported by NIH P41 RR-01081). This work was supported by an American Heart Association pre-doctoral fellowship (J.O'D) and the National Institutes of Health (R01 GM 66875, M.S.C.).

PY - 2009/3/15

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N2 - 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.

AB - 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.

KW - Electron microscopy

KW - Gene therapy

KW - Heparan sulfate

KW - Heparin

KW - Parvovirus

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Adeno-associated virus-2 and its primary cellular receptor-Cryo-EM structure of a heparin complex

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Keywords

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