Journal: J Virol / Year: 2007 Title: Structure of immature West Nile virus. Authors: Ying Zhang / Bärbel Kaufmann / Paul R Chipman / Richard J Kuhn / Michael G Rossmann / Abstract: The structure of immature West Nile virus particles, propagated in the presence of ammonium chloride to block virus maturation in the low-pH environment of the trans-Golgi network, was determined by ...The structure of immature West Nile virus particles, propagated in the presence of ammonium chloride to block virus maturation in the low-pH environment of the trans-Golgi network, was determined by cryo-electron microscopy (cryo-EM). The structure of these particles was similar to that of immature West Nile virus particles found as a minor component of mature virus samples (naturally occurring immature particles [NOIPs]). The structures of mature infectious flaviviruses are radically different from those of the immature particles. The similarity of the ammonium chloride-treated particles and NOIPs suggests either that the NOIPs have not undergone any conformational change during maturation or that the conformational change is reversible. Comparison with the cryo-EM reconstruction of immature dengue virus established the locations of the N-linked glycosylation sites of these viruses, verifying the interpretation of the reconstructions of the immature flaviviruses.
Electron dose: 22 e/Å2 / Film or detector model: KODAK SO-163 FILM
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Processing
EM software
ID
Name
Category
Details
1
EMfit
modelfitting
2
EM3DR
3Dreconstruction
3
PFT
3Dreconstruction
PFTSEARCH
4
POR
3Dreconstruction
CTF correction
Details: CTF correction of each particle
Symmetry
Point symmetry: I (icosahedral)
3D reconstruction
Method: PFT / Resolution: 24 Å / Num. of particles: 341 / Actual pixel size: 2.95 Å / Symmetry type: POINT
Atomic model building
Protocol: RIGID BODY FIT / Space: REAL Target criteria: best fit using criterial described in paper by Rossmann et al., Journal of structural biology 136, 190-200 (2001) Details: METHOD--Three E molecules were fitted consecutively REFINEMENT PROTOCOL--rigid body
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