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120 a.a.*
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269 a.a.*
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256 a.a.*
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238 a.a.*
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55 a.a.*
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* C-alpha coords only
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References listed in PDB file
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Key reference
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Title
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Interaction of coxsackievirus b3 with the full length coxsackievirus-Adenovirus receptor.
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Authors
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Y.He,
P.R.Chipman,
J.Howitt,
C.M.Bator,
M.A.Whitt,
T.S.Baker,
R.J.Kuhn,
C.W.Anderson,
P.Freimuth,
M.G.Rossmann.
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Ref.
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Nat Struct Biol, 2001,
8,
874-878.
[DOI no: ]
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PubMed id
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Abstract
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Group B coxsackieviruses (CVB) utilize the coxsackievirus-adenovirus receptor
(CAR) to recognize host cells. CAR is a membrane protein with two Ig-like
extracellular domains (D1 and D2), a transmembrane domain and a cytoplasmic
domain. The three-dimensional structure of coxsackievirus B3 (CVB3) in complex
with full length human CAR and also with the D1D2 fragment of CAR were
determined to approximately 22 A resolution using cryo-electron microscopy
(cryo-EM). Pairs of transmembrane domains of CAR associate with each other in a
detergent cloud that mimics a cellular plasma membrane. This is the first view
of a virus-receptor interaction at this resolution that includes the
transmembrane and cytoplasmic portion of the receptor. CAR binds with the distal
end of domain D1 in the canyon of CVB3, similar to how other receptor molecules
bind to entero- and rhinoviruses. The previously described interface of CAR with
the adenovirus knob protein utilizes a side surface of D1.
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Figure 2.
Figure 2. Orthogonal stereo views of the C  backbone
of CAR D1 and D2 (black) fit into the cryo-EM density.
Fragments of CVB3 (blue for VP1, green for VP2 and red for VP3)
are also shown. a, The south rim of the canyon, formed by VP2,
contacts the A and G  -strands
of domain D1. b, The sugar moieties of the carbohydrate at Asn
108 are depicted in yellow.
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Figure 3.
Figure 3. Stereo diagrams of the ICAM-1, PVR and CAR D1 domains.
The -strands
are labeled A -G. The amino acids identified as being in the
virus -receptor interface are indicated by spheres. a, ICAM-1
with HRV14 and HRV16 in blue, and ICAM-1 with coxsackievirus A21
in red; b, PVR with poliovirus in red; and c, CAR with
adenovirus knob in blue and CAR with CVB3 in red. d, Schematic
diagram of the modes by which CAR (green) binds to CVB3 (red)
and adenovirus19 (blue). The suggested membrane curvature is
speculative.
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The above figures are
reprinted
by permission from Macmillan Publishers Ltd:
Nat Struct Biol
(2001,
8,
874-878)
copyright 2001.
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Secondary reference #1
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Title
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Isolation of a common receptor for coxsackie b viruses and adenoviruses 2 and 5.
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Authors
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J.M.Bergelson,
J.A.Cunningham,
G.Droguett,
E.A.Kurt-Jones,
A.Krithivas,
J.S.Hong,
M.S.Horwitz,
R.L.Crowell,
R.W.Finberg.
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Ref.
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Science, 1997,
275,
1320-1323.
[DOI no: ]
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PubMed id
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Figure 3.
Fig. 3. Adenovirus interaction with CAR on transfected CHO
cells. (A through C) show 35S-labeled virus and fiber attachment
to transfected cells (30). Each panel shows mean values for
virus or fiber bound ± SD for triplicate monolayers and
is representative of at least two experiments. (A) Adenovirus
attachment. CHO-CAR and CHO-al 2 monolayers in 24-well plates
were incubated with labeled adenovirus 2 (Ad2; 20,000 cpm) (30)
for 1 hour at room temperature, then washed^ and dissolved for
scintillation counting. (B) Adenovirus fiber attachment.
Monolayers in six-well plates were incubated with labeled
adenovirus 2 fibers (25,000 cpm). (C) Inhibition of adenovirus
attachment by purified fibers and knob domains. CHO-CAR
monolayers were incubated with buffer (solid bar), isolated
adenovirus 2^ fibers (5 µg, hatched bar), or recombinant
adenovirus 5 knob domains (0.7 µg, stippled bar) before
addition of labeled adenovirus 2. (D) Scatchard analysis of
fiber binding. Duplicate CHO-CAR and HeLa monolayers were
incubated with 125I-labeled adenovirus 2 fibers at different
specific activities for 1 hour at 4°C. Monolayers were then
washed four times and^ bound radioactivity was determined.
Nonspecific binding was determined^ by incubating labeled fiber
in the presence of a 200-fold excess of unlabeled fiber.
Specific binding is shown as nanograms bound^ per million cells.
There was no specific binding to control CHO-al 2 monolayers.
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Figure 4.
Fig. 4. Adenovirus-mediated gene transfer. CHO-al 2 and
CHO-CAR cells in 24-well plates were exposed to Ad.CMV-  gal at
different multiplicities of infection [MOI (in PFU per cell)]
for 1 hour at room temperature, then unbound virus was removed
and cells were incubated for 40 hours at 37°C. Cells were
fixed with 2% paraformaldehyde and -Gal
activity was determined by incubation with phosphate-buffered
saline containing 5 mM ferric and 5 mM ferrous cyanide, 1 mM
MgCl[2], and X-Gal (1 mg/ml). Examination of monolayers before
staining revealed some cytotoxicity in the^ CHO-CAR cells
exposed to Ad.CMV- gal at 100
PFU per cell. This experiment was performed three times.
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The above figures are
reproduced from the cited reference
with permission from the AAAs
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Secondary reference #2
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Title
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Structural analysis of the mechanism of adenovirus binding to its human cellular receptor, Car.
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Authors
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M.C.Bewley,
K.Springer,
Y.B.Zhang,
P.Freimuth,
J.M.Flanagan.
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Ref.
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Science, 1999,
286,
1579-1583.
[DOI no: ]
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PubMed id
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Figure 2.
Fig. 2. A molecular surface representation of the interface in
the Ad12 knob-CAR D1 complex. (A) Sequence conservation surface
diagram of two knob monomers viewed at the CAR interface. The
molecules are colored on a sliding scale from white (conserved)
to red (nonconserved). Conservation analysis was based on an
alignment of all human Ad knob sequences available in GenBank. A
white strip of conservation transects the surface of the
molecule. Upon binding, the CAR D1 molecule occludes the
conserved strip on Ad12 knob. (B) Surface diagram of two
adjacent Ad12 knob monomers shown in the same view as (A). The
molecules are colored on a sliding scale from yellow (contact)
to red (no contact). Atoms in contact with CAR D1 are shared
between monomers. (C) Surface diagram of CAR D1. The molecules
are colored on a sliding scale from magenta (contact) to cyan
(no contact). This figure was generated with GRASP (30).
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Figure 3.
Fig. 3. CPK model of the region around the cavity. The three
consecutive proline residues in Ad12 knob partially shape the
cavity, which is colored magenta. The AB loop, whose carbon
atoms are colored yellow, lines one side of the cavity. The
carbon atoms from the remainder of the monomer are colored red,
those of the second knob monomer, green, and those of CAR D1,
cyan. All oxygen and nitrogen atoms are colored light red and
blue, respectively. The cavity is lined with atoms from residues
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 (backbone),
 (side),
V448 (side),  (backbone),
V450, L455 (side), Q535 (side), P573 (side), and S575 (side)
from one Ad12 knob; S514 (backbone), A515 (backbone),  (side),
N520 (side), A524 (main), E523, K525, and S526 (side) from the
other Ad12 knob; and L39 (side), K47 (backbone), V48 (backbone),
D49, Q50, V51, and K102 (side) from CAR. The underlined residues
are conserved or similar in all CAR-binding Ad serotypes.
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The above figures are
reproduced from the cited reference
with permission from the AAAs
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Secondary reference #3
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Title
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The structure of coxsackievirus b3 at 3.5 a resolution.
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Authors
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J.K.Muckelbauer,
M.Kremer,
I.Minor,
G.Diana,
F.J.Dutko,
J.Groarke,
D.C.Pevear,
M.G.Rossmann.
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Ref.
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Structure, 1995,
3,
653-667.
[DOI no: ]
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PubMed id
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Figure 1.
Figure 1. Stereo Cα traces for (a) VP1, (b) VP2, (c) VP3 and
(d) VP4 of CVB3 with every 20th residue labeled. Figure 1.
Stereo Cα traces for (a) VP1, (b) VP2, (c) VP3 and (d) VP4 of
CVB3 with every 20th residue labeled.
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Figure 5.
Figure 5. The water-accessible surfaces. Radial depth-cued
images viewed down the icosahedral twofold axis of (a) CVB3, (b)
PV1/M and (c) HRV14. Regions of dark blue color represent
surface depressions while regions of yellow color represent
surface protrusions. Figure 5. The water-accessible surfaces.
Radial depth-cued images viewed down the icosahedral twofold
axis of (a) CVB3, (b) PV1/M and (c) HRV14. Regions of dark blue
color represent surface depressions while regions of yellow
color represent surface protrusions. (Images courtesy of
Jean-Yves Sgro, Institute for Molecular Virology, University of
Wisconsin-Madison and created using the program GRASP [[3]78].)
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The above figures are
reproduced from the cited reference
with permission from Cell Press
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Headers
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