Scripps VIVO scripps research logo

  • Index
  • Log in
  • Home
  • People
  • Organizations
  • Research
  • Events
Search form

Cryoem structure at 9 Å resolution of an adenovirus vector targeted to hematopoietic cells

Academic Article
uri icon
  • Overview
  • Research
  • Identity
  • Additional Document Info
  • View All
scroll to property group menus

Overview

authors

  • Saban, S. D.
  • Nepomuceno, R. R.
  • Gritton, L. D.
  • Nemerow, Glen
  • Stewart, P. L.

publication date

  • June 2005

journal

  • Journal of Molecular Biology  Journal

abstract

  • We report a sub-nanometer resolution cryo-electron microscopy (cryoEM) structural analysis of an adenoviral vector, Ad35F, comprised of an adenovirus type 5 (Ad5) capsid pseudo-typed with an Ad35 fiber. This vector transduces human hematopoietic cells via association of its fiber protein with CD46, a member of the complement regulatory protein family. Major advances in data acquisition and image processing allowed a significant improvement in resolution compared to earlier structures. Analysis of the cryoEM density was enhanced by docking the crystal structures of both the hexon and penton base capsid proteins. CryoEM density was observed for hexon residues missing from the crystal structure that include hypervariable regions and the epitope of a neutralizing monoclonal antibody. Within the penton base, density was observed for the integrin-binding RGD loop missing from the crystal structure and for the flexible beta ribbon of the variable loop on the side of the penton base. The Ad35 fiber is flexible, consistent with the sequence insert in the third beta-spiral repeat. On the inner capsid surface density is revealed at the base of the hexons and below the penton base. A revised model is presented for protein IX within the virion. Well-defined density was assigned to a conserved domain in the N terminus of protein IX required for incorporation into the virion. For the C-terminal domain of protein IX two alternate conformations are proposed, either binding on the capsid surface or extending away from the capsid. This model is consistent with the tolerance of the C terminus for inserted ligands and its potential use in vector retargeting. This structural study increases our knowledge of Ad capsid assembly, antibody neutralization mechanisms, and may aid further improvements in gene delivery to important human cell types.

subject areas

  • Adenoviridae
  • Cryoelectron Microscopy
  • Gene Transfer Techniques
  • Genetic Vectors
  • Hematopoietic Stem Cells
  • Leukemia
  • Protein Structure, Tertiary
  • Viral Proteins
scroll to property group menus

Research

keywords

  • adenovirus
  • cryo-electron microscopy
  • cryoEM difference mapping
  • protein IX
  • single particle reconstruction
scroll to property group menus

Identity

International Standard Serial Number (ISSN)

  • 0022-2836

Digital Object Identifier (DOI)

  • 10.1016/j.jmb.2005.04.034

PubMed ID

  • 15890367
scroll to property group menus

Additional Document Info

start page

  • 526

end page

  • 537

volume

  • 349

issue

  • 3

©2021 The Scripps Research Institute | Terms of Use | Powered by VIVO

  • About
  • Contact Us
  • Support