3.3 | Dynamic stability of designed OsL1:OsS1 heterotetramers
To gain a better understanding of the crucial interfaces and residues implicated in the interaction between OsL1 and OsS1, we initially constructed three hypothetical models of OsL1:OsS1 heterotetramers (C-I, C-II, and C-III; Figure 3A). Subsequently, we performed molecular simulations for a cumulative duration of 100 ns (2 replicates) to investigate the dynamics of these complexes. After obtaining the MD trajectories, we analyzed the backbone RMSD and Rg of the complexes to gauge the complex stability and compactness. As shown in Figure 3, the RMSD of C-II conversed immediately after 3-5 ns of the production run (Figure 3C), while the C-I and C-III showed RMSD with significantly higher deviations (~6-8Å) (Figure 3B, 3D). Moreover, we observed a more stable and conserved gyradius in C-II compared to C-I and C-III (Figure 3E-3G). In addition, the conformational snapshots of pre- and post-MD snapshots of heterotetramer models (Figure S2) and intermolecular H-bonds at different interfaces with respect to the simulation time (Figure S3) indicated a stable association of the OsL1 and OsS1 subunits in C-II.