Figure 3. Stabilization Strategies for Pre-Fusion LASV Glycoprotein Conformation.

(A) GP1-GP2 interface highlighted for stabilization, showing the α5 helix (from GP1) interaction with α6 and α7 helices (from GP2 subunit). (B) Residues (K320, R325, K327, E329) from the post-fusion 6HB conformational ensemble that form stabilizing interactions, illustrating their importance in pre-fusion stability. (C) Overview of the ProteinMPNN design protocol applied to the pre-fusion LASV GPC structure (PDB: 7PVD), showcasing the redesign strategy to enhance structural stability while preserving mAb binding epitopes and N-linked glycosylation sites. The generation of 2,306 sequences from varying temperature parameters and models is illustrated, leading to a refinement process that identified the top 24 designs for experimental evaluation.