Interaction of Neurotropic Enterovirus-71 with Receptor PSGL-1
Matthew Makowski ¹, Josh Yoder ², Javier Cifuente ², Bob Ashley ², Jennifer Yoder ², Hyunwook Lee ², James Conway ³,
Yorihiro Nishimura ⁴, Susan Hafenstein ²
¹ American University, Washington, DC, ² Penn State College of Medicine, Department of Medicine and Department of Microbiology and
Immunology, Hershey, PA, ³ University of Pittsburgh, Pittsburgh, PA, ⁴ National Institute of Infectious Diseases, Tokyo, Japan
Results
Introduction
Enterovirus 71 (EV71) is a picornavirus
with pandemic potential. EV71 infection is
a major causative agent of hand, foot, and
mouth disease (HFMD), typically a mild,
self-limiting disease that can result in
severe neurological symptoms and
significant pediatric mortality. Identification
of viral receptors is crucial to
understanding the infection mechanism of
EV71 in the search for potential antiviral
targets. We performed a single-particle
cyro-electron microscopy reconstruction of
EV71 with bound P-selectin glycoprotein
ligand-1 (psgl-1), previously identified as a
functional receptor for EV71, to indicate
candidate viral binding sites for the psgl-1
cellular receptor.
Methods
Figure 5: Difference map analysis of EV71+psgl-1 compared with EV71 native particle
Left: EV71+psgl-1 reconstruction, 1,932 particles, 15.89 A resolution
Center: Native EV71 particle (no bound psgl-1) at approximately 10 A
resolution.
Figure 1: Structure of
truncated psgl-1 receptor
Figure 3: Negatively stained electron micrograph images of
EV71 with bound psgl-1
Right: Difference map displaying density differences between psgl-1-bound
and psgl-1-unbound virus reconstructions. Significant density is apparent
surrounding the icosahedral five-fold and three-fold vertices.
Figure 7: Native EV71/EV71+psgl-1 difference map superimposed with BEV polyprotein
EV71 VP1-145 (necessary for EV71 infectivity in psgl-1
expressing cell lines) correlated with BEV VP1-131
(shown as red spheres), shown in close proximity to
regions of high difference density near the five-fold vertex
indicating a potential binding site for psgl-1.
Acknowledgments
-NIH Grant K22 AI079271-02 Grant
-2011 C. Max Lang Junior Faculty Research Scholar Award
-Hershey Medical Center Core Imaging Facility
-University of Pittsburgh Department of Structural Biology Imaging
Facility
-Hershey Medical Center Summer Undergraduate Research Internship
Program
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Figure 4: Structural comparison of EV71 using three different data sets.
VP1-145
Left: EV71+psgl-1 reconstruction performed using 1,614 boxed micrograph images to
an estimated resolution of 18.13 A.
Figure 2: General method of performing computer-aided viral reconstructions
Figure 6: Sequence alignment for EV71 VP1 and Bovine Enterovirus (BEV) VP1,
which has a known x-ray structure.
Bovine enterovirus (BEV) displays the highest
Center: EV71+psgl-1 reconstruction performed using 658 boxed micrograph images to sequence homology of viruses with known
an estimated resolution of 15.38 A.
crystal structures, so BEV can be used
analogously to study putative binding properties
Right: EV71+psgl-1 reconstruction performed using 85% of the combined data set
of EV71 with psgl-1.
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