Supplementary Materials

A glycerophospholipid-specific pocket in the RVFV class II fusion protein drives target membrane insertion

Pérez-Vargas, S. M. de Boer, M. A. Tortorici, G. Pehau-Arnaudet, J. Lepault, P. England, P. J. Rottier, B. J. Bosch, J. S. Hub, F. A. Rey

Materials/Methods, Supplementary Text, Tables, Figures, and/or References

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  • Materials and Methods 
  • Figs. S1 to S8 
  • Tables S1 to S3 
  • Captions for Movies S1 to S5 
  • References

Images, Video, and Other Media

Movie S1
Superposition of the Gc subunits in the various conformations. The Gc subunit in the monomer (pdb 4HJ1), dimer (pdb 4HJC) (both from (8)) and trimer (this work), are shown in ribbons colored according to the key on the side and superposed on domain II and most of domain I, showing that there is no difference in hinge angle between domains I and II as in other class-II fusion proteins, and that the conformational change is confined to a large rearrangement of domain III about the linker between domains I and III, and to the region of domain I from which the linker extends (i.e., the end of domain I that is opposite to the domain Iô€,±II interface). The invariable parts of the Gc subunit superpose with an rmsd of 0.9 Å over 326 atoms (monomer vs dimer), 1.55 Å over 283 atoms (trimer vs dimer) and 2.23 Å over 295 atoms (monomer vs trimer).
Movie S2
Morph of the transition from pre-fusion to post-fusion form. The two structures reported previously of the pre-fusion form (dimer, pdb 4HJ1; and monomer, pdb 4HJC) were used. As proposed by Dessau and Modis, the structure of the monomer appears as an extended intermediate, and it was not possible to morph directly from the dimer subunit (4HJ1) to that of the post-fusion trimer without introducing serious clashes. Instead, morphing first from the conformation of Gc in the dimer to that observed in the monomer (4HJC), allows to then transit to the final post-fusion form in the trimer without clashing.
Movie S3
Morph of the transition from pre-fusion to post-fusion form, as in Movie S2, showing that the side chains of Phe866 and Phe1018 make part of the hydrophobic core of domain I in the pre-fusion forms, and are dislodged from that location to become part of the inter-subunit interface within the post-fusion trimer. Phe1022, in the linker, also undergoes an important relocation.
Movie S4
Quality of the electron density map for C3PC binding pocket. Electron density maps colored at 1ô€�- around the protein (blue map) and 0.9ô€�- around the C3PC molecule (wheat map). The residues involved in binding are represented as sticks and labeled.
Movie S5
Molecular dynamics simulations of the interactions of Gc with membranes. Segments with duration of 150 ns from three different simulations are shown, with membranes containing only DOPC (left), DOPC plus 20% cholesterol (central) and 40% cholesterol (right). The polar heads of DOPC are represented as spheres and the aliphatic moiety as gray lines, the cholesterol is shown as red lines. The protein is shown in cartoon representation with the residues involved in the C3PC binding pocket as green spheres.