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Proc Natl Acad Sci U S A. 2014 Mar 18;111(11):4285-90. doi: 10.1073/pnas.1322036111. Epub 2014 Mar 4.

Structural basis of Flavivirus NS1 assembly and antibody recognition.

Author information

1
Departments of Pathology and Immunology, Medicine, Molecular Microbiology, and Biochemistry and Molecular Biophysics, Washington University in St. Louis, St. Louis, MO 63110.

Abstract

The Flavivirus nonstructural protein 1 (NS1) is a conserved, membrane-associated and secreted glycoprotein with replication and immune evasion functions. Secreted NS1 is a hexameric, barrel-shaped lipoprotein that can bind back to the plasma membrane of cells. Antibodies targeting cell surface-associated NS1 can be protective in vivo in a manner dependent on Fc effector functions. We describe here the crystal structure of a C-terminal fragment (residues 172-352) of West Nile (WNV) and Dengue virus NS1 proteins at 1.85 and 2.7 Å resolution, respectively. NS1(172-352) assembles as a unique rod-shaped dimer composed of a 16-stranded β-platform flanked on one face by protruding connecting loops. We also determined the 3.0 Å resolution structure of WNV NS1(172-352) with the protective 22NS1 antibody Fab, which engages the loop-face of the rod. The head-to-head NS1(172-352) dimer we observe in crystal lattices is supported by multiangle light and small-angle X-ray scattering studies. We used the available cryo-electron microscopy reconstruction to develop a pseudoatomic model of the NS1 hexamer. The model was constructed with the NS1(172-352) dimeric rod aligned with the long axis of the barrel, and with the loop-face oriented away from the core. Difference densities suggest that the N-terminal region of NS1 forms globular lobes that mediate lateral contacts between dimers in the hexamer. Our model also suggests that the N-terminal lobe forms the surface of the central cavity where lipid binding may occur.

KEYWORDS:

human pathogen; immune epitope; structural biology; virology

PMID:
24594604
PMCID:
PMC3964132
DOI:
10.1073/pnas.1322036111
[Indexed for MEDLINE]
Free PMC Article

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