Capturing a flavivirus pre-fusion intermediate.
During cell entry of flaviviruses, low endosomal pH triggers the rearrangement of the viral surface glycoproteins to a fusion-active state that allows the release of the infectious RNA into the cytoplasm. In this work, West Nile virus was complexed with Fab fragments of the neutralizing mAb E16 and was subsequently exposed to low pH, trapping the virions in a pre-fusion intermediate state. The structure of the complex was studied by cryo-electron microscopy and provides the first structural glimpse of a flavivirus fusion intermediate near physiological conditions. A radial expansion of the outer protein layer of the virion was observed compared to the structure at pH 8. The resulting approximately 60 A-wide shell of low density between lipid bilayer and outer protein layer is likely traversed by the stem region of the E glycoprotein. By using antibody fragments, we have captured a structural intermediate of a virus that likely occurs during cell entry. The trapping of structural transition states by antibody fragments will be applicable for other processes in the flavivirus life cycle and delineating other cellular events that involve conformational rearrangements.
Kaufmann
Bärbel
B
Department of Biological Sciences, Purdue University, West Lafayette, Indiana, United States of America.
Chipman
Paul R
PR
Holdaway
Heather A
HA
Johnson
Syd
S
Fremont
Daved H
DH
Kuhn
Richard J
RJ
Diamond
Michael S
MS
Rossmann
Michael G
MG
D000914:Antibodies, Viral; D020285:Cryoelectron Microscopy; D005416:Flavivirus / Q000737:chemistry / Q000472:pathogenicity*; D006023:Glycoproteins; D006863:Hydrogen-Ion Concentration; D007128:Immunoglobulin Fragments / Q000378:metabolism; D008051:Lipid Bilayers; D014759:Viral Envelope Proteins / Q000737:chemistry; D014760:Viral Fusion Proteins / Q000737:chemistry*; D014771:Virion / Q000737:chemistry; D053586:Virus Internalization
D000914:Antibodies, Viral; D006023:Glycoproteins; D007128:Immunoglobulin Fragments; D008051:Lipid Bilayers; D014759:Viral Envelope Proteins; D014760:Viral Fusion Proteins