The 2.8 Å Electron Microscopy Structure of Adeno-Associated Virus-DJ Bound by a Heparinoid Pentasaccharide
Atomic structures of adeno-associated virus (AAV)-DJ, alone and in complex with fondaparinux, have been determined by cryoelectron microscopy at 3 Å resolution. The gene therapy vector, AAV-DJ, is a hybrid of natural serotypes that was previously derived by directed evolution, selecting for hepatocy...
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| Format: | Article |
| Language: | English |
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Elsevier
2017-06-01
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| Series: | Molecular Therapy: Methods & Clinical Development |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S232905011730044X |
| _version_ | 1828794271664177152 |
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| author | Qing Xie John M. Spear Alex J. Noble Duncan R. Sousa Nancy L. Meyer Omar Davulcu Fuming Zhang Robert J. Linhardt Scott M. Stagg Michael S. Chapman |
| author_facet | Qing Xie John M. Spear Alex J. Noble Duncan R. Sousa Nancy L. Meyer Omar Davulcu Fuming Zhang Robert J. Linhardt Scott M. Stagg Michael S. Chapman |
| author_sort | Qing Xie |
| collection | DOAJ |
| description | Atomic structures of adeno-associated virus (AAV)-DJ, alone and in complex with fondaparinux, have been determined by cryoelectron microscopy at 3 Å resolution. The gene therapy vector, AAV-DJ, is a hybrid of natural serotypes that was previously derived by directed evolution, selecting for hepatocyte entry and resistance to neutralization by human serum. The structure of AAV-DJ differs from that of parental serotypes in two regions where neutralizing antibodies bind, so immune escape appears to have been the primary driver of AAV-DJ’s directed evolution. Fondaparinux is an analog of cell surface heparan sulfate to which several AAVs bind during entry. Fondaparinux interacts with viral arginines at a known heparin binding site, without the large conformational changes whose presence was controversial in low-resolution imaging of AAV2-heparin complexes. The glycan density suggests multi-modal binding that could accommodate sequence variation and multivalent binding along a glycan polymer, consistent with a role in attachment, prior to more specific interactions with a receptor protein mediating entry. |
| first_indexed | 2024-12-12T03:41:10Z |
| format | Article |
| id | doaj.art-fb211d03182c45d4b321803376200cb3 |
| institution | Directory Open Access Journal |
| issn | 2329-0501 |
| language | English |
| last_indexed | 2024-12-12T03:41:10Z |
| publishDate | 2017-06-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Molecular Therapy: Methods & Clinical Development |
| spelling | doaj.art-fb211d03182c45d4b321803376200cb32022-12-22T00:39:40ZengElsevierMolecular Therapy: Methods & Clinical Development2329-05012017-06-015C11210.1016/j.omtm.2017.02.004The 2.8 Å Electron Microscopy Structure of Adeno-Associated Virus-DJ Bound by a Heparinoid PentasaccharideQing Xie0John M. Spear1Alex J. Noble2Duncan R. Sousa3Nancy L. Meyer4Omar Davulcu5Fuming Zhang6Robert J. Linhardt7Scott M. Stagg8Michael S. Chapman9Department of Biochemistry & Molecular Biology, School of Medicine, Oregon Health & Science University, Portland, OR 97239-3098, USAInstitute of Molecular Biophysics, Florida State University, 91 Chieftan Way, Tallahassee, FL 32306-4380, USAInstitute of Molecular Biophysics, Florida State University, 91 Chieftan Way, Tallahassee, FL 32306-4380, USAInstitute of Molecular Biophysics, Florida State University, 91 Chieftan Way, Tallahassee, FL 32306-4380, USADepartment of Biochemistry & Molecular Biology, School of Medicine, Oregon Health & Science University, Portland, OR 97239-3098, USADepartment of Biochemistry & Molecular Biology, School of Medicine, Oregon Health & Science University, Portland, OR 97239-3098, USADepartments of Chemical and Biological Engineering, Chemistry, and Chemical Biology, Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy, NY 12180, USADepartments of Chemical and Biological Engineering, Chemistry, and Chemical Biology, Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy, NY 12180, USAInstitute of Molecular Biophysics, Florida State University, 91 Chieftan Way, Tallahassee, FL 32306-4380, USADepartment of Biochemistry & Molecular Biology, School of Medicine, Oregon Health & Science University, Portland, OR 97239-3098, USAAtomic structures of adeno-associated virus (AAV)-DJ, alone and in complex with fondaparinux, have been determined by cryoelectron microscopy at 3 Å resolution. The gene therapy vector, AAV-DJ, is a hybrid of natural serotypes that was previously derived by directed evolution, selecting for hepatocyte entry and resistance to neutralization by human serum. The structure of AAV-DJ differs from that of parental serotypes in two regions where neutralizing antibodies bind, so immune escape appears to have been the primary driver of AAV-DJ’s directed evolution. Fondaparinux is an analog of cell surface heparan sulfate to which several AAVs bind during entry. Fondaparinux interacts with viral arginines at a known heparin binding site, without the large conformational changes whose presence was controversial in low-resolution imaging of AAV2-heparin complexes. The glycan density suggests multi-modal binding that could accommodate sequence variation and multivalent binding along a glycan polymer, consistent with a role in attachment, prior to more specific interactions with a receptor protein mediating entry.http://www.sciencedirect.com/science/article/pii/S232905011730044XAAVadeno-associated virusglycanheparan sulfateattachmentreceptorstructureelectron microscopygene therapyvector |
| spellingShingle | Qing Xie John M. Spear Alex J. Noble Duncan R. Sousa Nancy L. Meyer Omar Davulcu Fuming Zhang Robert J. Linhardt Scott M. Stagg Michael S. Chapman The 2.8 Å Electron Microscopy Structure of Adeno-Associated Virus-DJ Bound by a Heparinoid Pentasaccharide Molecular Therapy: Methods & Clinical Development AAV adeno-associated virus glycan heparan sulfate attachment receptor structure electron microscopy gene therapy vector |
| title | The 2.8 Å Electron Microscopy Structure of Adeno-Associated Virus-DJ Bound by a Heparinoid Pentasaccharide |
| title_full | The 2.8 Å Electron Microscopy Structure of Adeno-Associated Virus-DJ Bound by a Heparinoid Pentasaccharide |
| title_fullStr | The 2.8 Å Electron Microscopy Structure of Adeno-Associated Virus-DJ Bound by a Heparinoid Pentasaccharide |
| title_full_unstemmed | The 2.8 Å Electron Microscopy Structure of Adeno-Associated Virus-DJ Bound by a Heparinoid Pentasaccharide |
| title_short | The 2.8 Å Electron Microscopy Structure of Adeno-Associated Virus-DJ Bound by a Heparinoid Pentasaccharide |
| title_sort | 2 8 a electron microscopy structure of adeno associated virus dj bound by a heparinoid pentasaccharide |
| topic | AAV adeno-associated virus glycan heparan sulfate attachment receptor structure electron microscopy gene therapy vector |
| url | http://www.sciencedirect.com/science/article/pii/S232905011730044X |
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