http://www.plosntds.org/article/info:doi/10.1371/journal.pntd.0000721
Abstract
Viral fusogenic envelope proteins are important targets for the development of inhibitors of viral entry. We report an approach for the computational design of peptide inhibitors of the dengue 2 virus (DENV-2) envelope (E) protein using high-resolution structural data from a pre-entry dimeric form of the protein. By using predictive strategies together with computational optimization of binding ‘‘pseudoenergies’’, we were able to design multiple peptide sequences that showed low micromolar viral entry inhibitory activity. The two most active peptides, DN57opt and 1OAN1, were designed to displace regions in the domain II hinge, and the first domain I/domain II beta sheet connection, respectively, and show fifty percent inhibitory concentrations of 8 and 7 mM respectively in a focus forming unit assay. The antiviral peptides were shown to interfere with virus:cell binding, interact directly with the E proteins and also cause changes to the viral surface using biolayer interferometry and cryo-electron microscopy, respectively. These peptides may be useful for characterization of intermediate states in the membrane fusion process, investigation of DENV receptor molecules, and as lead compounds
for drug discovery.
See Figure 5.
Peptide:E protein binding assay. Biolayer interferometry was used to assay the binding of the peptides to truncated E Protein. The
association and dissociation of increasing concentrations of truncated E protein to peptides DN57opt (A) and 1OAN1 (B) are shown. A buffer blank
(PBS, 0.02% Tween-20, 0.1% BSA) containing no E protein was run for each peptide. The affinity of the peptides for the truncated E protein was
calculated (DN57opt KD = 1.26102660.661026 M (mean6sd), 1OAN1 KD = 4.56102762.061027 M).
doi:10.1371/journal.pntd.0000721.g005
We thank the authors below for their contribution.
Joshua M. Costin1, Ekachai Jenwitheesuk2¤a, Shee-Mei Lok3¤b, Elizabeth Hunsperger4, Kelly A. Conrads5, Krystal A. Fontaine1¤c, Craig R. Rees1¤d, Michael G. Rossmann3, Sharon Isern1, Ram Samudrala2*, Scott F. Michael1*
1 Department of Biological Sciences, Florida Gulf Coast University, Fort Myers, Florida, United States of America, 2 Department of Microbiology, University of Washington, Seattle, Washington, United States of America, 3 Department of Biological Sciences, Purdue University, West Lafayette, Indiana, United States of America, 4 Dengue Branch, Division of Vector-Borne Infectious Diseases, Centers for Disease Control and Prevention, San Juan, Puerto Rico, 5 Forte´Bio, Incorporated, Menlo Park, California, United States of America
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