Structural Studies of Coronavirus Fusion Proteins
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Principal Investigator: David Veesler Organization: UNIVERSITY OF WASHINGTON Fiscal Year: 2019 Award: $293,325 Funding agency: National Institute of General Medical Sciences Enveloped viruses use specialized proteins present at the virus surface to translocate their genetic material across the host cell membrane during infection. For coronaviruses, homotrimers of the spike glycoprotein promote host cell attachment and fusion of the viral and host membranes. Although coronaviruses have a significant pandemic potential, the lack of high-resolution data for any coronavirus spike trimer limits our mechanistic understanding of infection by this family of viruses. The objective of the proposed work is to obtain high-resolution snapshots corresponding to the various stages of the fusion reaction mediated by coronavirus spikes and to study the structural determinants associated with antibody inhibition of viral infection. In Aim I, we propose to elucidate the architecture of the Mouse Hepatitis Virus (MHV) pre-fusion spike using cryoEM. Aim II will be dedicated to studying the conformational changes associated with the fusion reaction with an emphasis on the first intermediate (extended intermediate) and the post-fusion spike. In Aim III, we will characterize the 3D organization of human coronavirus spikes to understand how these viruses overcome the species barrier and to identify structurally conserved regions that could be potential targets for therapeutic initiatives. The final aim will rely on structure-guided protein design to engineer antibodies targeting human coronavirus spikes with the goal of identifying immunogens for raising broadly-neutralizing antibodies. Terms: <3-D><3-D modeling><3-Dimensional><3D><3D modeling><ATGN><Address><Antibodies><Antibody Fragments><Antigens><Architecture><Assay><Attention><Binding><Bioassay><Biologic Assays><Biologic Models><Biological Assay><Biological Models><Cell Attachment><Cell Body><Cell fusion><Cell membrane><Cell-Matrix Adhesions><Cell-Matrix Junction><Cells><Chimera Protein><Chimeric Proteins><CoV spike glycoprotein><Complement><Complement Proteins><Complex><Coronaviridae><Coronaviridae Infections><Coronavirus><Coronavirus Infections><Coronavirus spike protein><Cryo-electron Microscopy><Cryoelectron Microscopy><Cytoplasmic Membrane><Data><Data Analyses><Data Analysis><Disease Outbreaks><Electron Cryomicroscopy><Engineering / Architecture><Family><Fostering><Fusion Protein><Future><General Viruses><Genetic Materials><Genetics-Mutagenesis><Genome><Glycoproteins><Goals><Grips><Health><Human><Immunization><Immunoglobulin Fragments><Immunologic Sensitization><Immunologic Stimulation><Immunological Sensitization><Immunological Stimulation><Immunostimulation><Infection><Kinetics><Lead><MERS-CoV><Mediating><Membrane><Membrane Fusion><Mice><Mice Mammals><Middle East Respiratory Syndrome><Middle East Respiratory Syndrome Coronavirus><Model System><Modeling><Modern Man><Molecular><Molecular Configuration><Molecular Conformation><Molecular Interaction><Molecular Stereochemistry><Mouse Hepatitis Virus><Murine><Murine Gastroenteritis Virus><Murine hepatitis virus><Mus><Mutagenesis><Mutagenesis Molecular Biology><N-terminal><NH2-terminal><Negative Staining><Organization Charts><Outbreaks><Outcome><Pathogenicity><Pb element><Plasma Membrane><Pneumonia><Process><Protein Engineering><Proteins><Reaction><Receptor Protein><Resolution><Structure><Surface><Thermodynamic><Thermodynamics><Viral><Viral Diseases><Viral Fusion Proteins><Virus><Virus Diseases><Work><antibody engineering><antibody inhibitor><biophysical approaches><biophysical methodology><biophysical methods><biophysical techniques><conformation><conformational state><coronavirus spike glycoprotein><cryo-EM><cryoEM><data interpretation><design><designing><experiment><experimental research><experimental study><genetic protein engineering><grasp><heavy metal Pb><heavy metal lead><immunogen><membrane structure><mutant><neutralizing antibody><organizational structure><pandemic><pandemic disease><particle><plasmalemma><prevent><preventing><protein design><receptor><receptor binding><receptor bound><reconstruction><small molecule inhibitor><stem><targeted drug therapy><targeted drug treatments><targeted therapeutic><targeted therapeutic agents><targeted therapy><targeted treatment><therapeutic target><three-dimensional modeling><vaccinology><viral infection><virus envelope><virus infection><virus-induced disease>