Journal: EMBO J / Year: 2003 Title: Structures of immature flavivirus particles. Authors: Ying Zhang / Jeroen Corver / Paul R Chipman / Wei Zhang / Sergei V Pletnev / Dagmar Sedlak / Timothy S Baker / James H Strauss / Richard J Kuhn / Michael G Rossmann / Abstract: Structures of prM-containing dengue and yellow fever virus particles were determined to 16 and 25 A resolution, respectively, by cryoelectron microscopy and image reconstruction techniques. The ...Structures of prM-containing dengue and yellow fever virus particles were determined to 16 and 25 A resolution, respectively, by cryoelectron microscopy and image reconstruction techniques. The closely similar structures show 60 icosahedrally organized trimeric spikes on the particle surface. Each spike consists of three prM:E heterodimers, where E is an envelope glycoprotein and prM is the precursor to the membrane protein M. The pre-peptide components of the prM proteins in each spike cover the fusion peptides at the distal ends of the E glycoproteins in a manner similar to the organization of the glycoproteins in the alphavirus spikes. Each heterodimer is associated with an E and a prM transmembrane density. These transmembrane densities represent either an EE or prMprM antiparallel coiled coil by which each protein spans the membrane twice, leaving the C-terminus of each protein on the exterior of the viral membrane, consistent with the predicted membrane-spanning domains of the unprocessed polyprotein.
SEQUENCE AUTHORS PROVIDED COORDINATES FOR ALPHA CARBONS ONLY. THE SEQRES RECORDS ARE FROM TICK-BORN ...SEQUENCE AUTHORS PROVIDED COORDINATES FOR ALPHA CARBONS ONLY. THE SEQRES RECORDS ARE FROM TICK-BORN ENCEPHALITIS VIRUS BECAUSE THE FITTING MODEL OF MAJOR ENVELOPE PROTEIN E (1SVB) WAS USED IN THIS STUDY.
A: major envelope protein E B: major envelope protein E C: major envelope protein E
x 5
icosahedral pentamer
648 kDa, 15 polymers
Theoretical mass
Number of molelcules
Total (without water)
648,467
15
Polymers
648,467
15
Non-polymers
0
0
Water
0
Type
Name
Symmetry operation
Number
identity operation
1_555
x,y,z
1
point symmetry operation
4
4
A: major envelope protein E B: major envelope protein E C: major envelope protein E
x 6
icosahedral 23 hexamer
778 kDa, 18 polymers
Theoretical mass
Number of molelcules
Total (without water)
778,161
18
Polymers
778,161
18
Non-polymers
0
0
Water
0
Type
Name
Symmetry operation
Number
identity operation
1_555
x,y,z
1
point symmetry operation
5
5
Idetical with deposited unit in distinct coordinate
icosahedral asymmetric unit, std point frame
Type
Name
Symmetry operation
Number
transform to point frame
1
Symmetry
Point symmetry: (Hermann–Mauguin notation: 532 / Schoenflies symbol: I (icosahedral))
-
Components
#1: Protein
majorenvelopeproteinE / Coordinate model: Cα atoms only
Mass: 43231.145 Da / Num. of mol.: 3 / Source method: isolated from a natural source / Source: (natural) Yellow fever virus / Genus: Flavivirus / References: UniProt: P14336
-
Experimental details
-
Experiment
Experiment
Method: ELECTRON MICROSCOPY
EM experiment
Aggregation state: PARTICLE / 3D reconstruction method: single particle reconstruction
-
Sample preparation
Component
Name: Yellow fever immature virus / Type: VIRUS Details: The samples were produced by adding ammonium chloride to the medium in the late infection stage.
Specimen
Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES
Vitrification
Cryogen name: ETHANE
-
Electron microscopy imaging
Microscopy
Model: FEI/PHILIPS CM300FEG/T
Electron gun
Electron source: FIELD EMISSION GUN / Illumination mode: FLOOD BEAM
Electron lens
Mode: BRIGHT FIELD
Image recording
Electron dose: 15 e/Å2 / Film or detector model: KODAK SO-163 FILM
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