4R0R

Ebolavirus GP Prehairpin Intermediate Mimic

Summary for 4R0R

• Entry DOI: 10.2210/pdb4r0r/pdb
• Related: 1EBO 2EBO
• Descriptor: eboIZN21 (2 entities in total)
• Functional Keywords: coiled-coil, n-trimer, prehairpin intermediate, biosynthetic protein
• Biological source: synthetic construct
• Total number of polymer chains: 1
• Total formula weight: 5621.55

Authors

Clinton, T.R.,Weinstock, M.T.,Jacobsen, M.T.,Szabo-Fresnais, N.,Pandya, M.J.,Whitby, F.G.,Herbert, A.S.,Prugar, L.I.,McKinnon, R.,Hill, C.P.,Welch, B.D.,Dye, J.M.,Eckert, D.M.,Kay, M.S. (deposition date: 2014-08-01, release date: 2014-10-22, Last modification date: 2024-11-06)

Primary citation

Clinton, T.R.,Weinstock, M.T.,Jacobsen, M.T.,Szabo-Fresnais, N.,Pandya, M.J.,Whitby, F.G.,Herbert, A.S.,Prugar, L.I.,McKinnon, R.,Hill, C.P.,Welch, B.D.,Dye, J.M.,Eckert, D.M.,Kay, M.S.
Design and characterization of ebolavirus GP prehairpin intermediate mimics as drug targets.
Protein Sci., 24:446-463, 2015

PubMed Abstract: Ebolaviruses are highly lethal filoviruses that cause hemorrhagic fever in humans and nonhuman primates. With no approved treatments or preventatives, the development of an anti-ebolavirus therapy to protect against natural infections and potential weaponization is an urgent global health need. Here, we describe the design, biophysical characterization, and validation of peptide mimics of the ebolavirus N-trimer, a highly conserved region of the GP2 fusion protein, to be used as targets to develop broad-spectrum inhibitors of ebolavirus entry. The N-trimer region of GP2 is 90% identical across all ebolavirus species and forms a critical part of the prehairpin intermediate that is exposed during viral entry. Specifically, we fused designed coiled coils to the N-trimer to present it as a soluble trimeric coiled coil as it appears during membrane fusion. Circular dichroism, sedimentation equilibrium, and X-ray crystallography analyses reveal the helical, trimeric structure of the designed N-trimer mimic targets. Surface plasmon resonance studies validate that the N-trimer mimic binds its native ligand, the C-peptide region of GP2. The longest N-trimer mimic also inhibits virus entry, thereby confirming binding of the C-peptide region during viral entry and the presence of a vulnerable prehairpin intermediate. Using phage display as a model system, we validate the suitability of the N-trimer mimics as drug screening targets. Finally, we describe the foundational work to use the N-trimer mimics as targets in mirror-image phage display, which will be used to identify D-peptide inhibitors of ebolavirus entry.

PubMed: 25287718
DOI: 10.1002/pro.2578

Experimental method

X-RAY DIFFRACTION (2.15 Å)