Construct designs
| Construct name | Fusion | Mutations | Cross-link |
| WT GCN4-Kar3Vik1 | GCN4 | NA | |
| Cysless GCN4-Kar3Vik1 | GCN4 | Kar3 (C391L, C469A, C517A, C566V, C655V)Vik1 (C377V, C436A, C536A, C596A, C640A) | NA |
| 10 Å cross-linked GCN4-Kar3Vik1 | GCN4 | Kar3 (C391L, C469A, C517A, C566V, C655V, E382C)Vik1 (C377V, C436A, C536A, C596A,C640A, K372C) | M4M |
| 20 Å cross-linked GCN4-Kar3Vik1 | GCN4 | Kar3 (C391L, C469A, C517A, C566V, C655V, E382C)Vik1 (C377V, C436A, C536A, C596A,C640A, K372C) | DPDPB |
| Non-cross-linked GCN4-Kar3Vik1 | GCN4 | Kar3 (C391L, C469A, C517A, C566V, C655V, E382C)Vik1 (C377V, C436A, C536A, C596A,C640A, K372C) | NA |
| WT SHD-Kar3Vik1 | SHD | NA | |
| Cysless SHD-Kar3Vik1 | SHD | Kar3 (C391L, C469A, C517A, C566V, C655V)Vik1 (C377V, C436A, C536A, C596A, C640A) | NA |
| Non-cross-linked SHD-Kar3Vik1 | SHD | Kar3 (C391L, C469A, C517A, C566V, C655V)Vik1 (C377V, C436A, C536A, C596A, C640A, E355C, K423C) | NA |
| Cross-linked SHD-Kar3Vik1 | SHD | Kar3 (C391L, C469A, C517A, C566V, C655V)Vik1 (C377V, C436A, C536A, C596A, C640A, E355C, K423C) | EBI |
| Construct name | Fusion | Mutations | Cross-link |
| WT GCN4-Kar3Vik1 | GCN4 | NA | |
| Cysless GCN4-Kar3Vik1 | GCN4 | Kar3 (C391L, C469A, C517A, C566V, C655V)Vik1 (C377V, C436A, C536A, C596A, C640A) | NA |
| 10 Å cross-linked GCN4-Kar3Vik1 | GCN4 | Kar3 (C391L, C469A, C517A, C566V, C655V, E382C)Vik1 (C377V, C436A, C536A, C596A,C640A, K372C) | M4M |
| 20 Å cross-linked GCN4-Kar3Vik1 | GCN4 | Kar3 (C391L, C469A, C517A, C566V, C655V, E382C)Vik1 (C377V, C436A, C536A, C596A,C640A, K372C) | DPDPB |
| Non-cross-linked GCN4-Kar3Vik1 | GCN4 | Kar3 (C391L, C469A, C517A, C566V, C655V, E382C)Vik1 (C377V, C436A, C536A, C596A,C640A, K372C) | NA |
| WT SHD-Kar3Vik1 | SHD | NA | |
| Cysless SHD-Kar3Vik1 | SHD | Kar3 (C391L, C469A, C517A, C566V, C655V)Vik1 (C377V, C436A, C536A, C596A, C640A) | NA |
| Non-cross-linked SHD-Kar3Vik1 | SHD | Kar3 (C391L, C469A, C517A, C566V, C655V)Vik1 (C377V, C436A, C536A, C596A, C640A, E355C, K423C) | NA |
| Cross-linked SHD-Kar3Vik1 | SHD | Kar3 (C391L, C469A, C517A, C566V, C655V)Vik1 (C377V, C436A, C536A, C596A, C640A, E355C, K423C) | EBI |
A list of the composition of all constructs including the identity of the stabilizing coiled coil, mutations, and cross-links. Kar3 constructs included a Tobacco etch virus (TEV) protease cleavable His-tag with the sequence: MGSSHHHHHHHHDYDIPTSENLYFQGASM followed by a fusion domain and K353-K729 of Kar3. Vik1 constructs included a TEV protease cleavable Strep tag with the sequence MASWSHPQFEKENLYFQGAS followed by a fusion domain and S341-T647 of Vik1. Amino acids in italics were removed by TEV protease. The sequences of GCN4, basic helix, and acidic helix in the fusions were: SVKELEDKVEELLSKNYHLENEVARLKKLV, KIAALKEKIAALKEKIAALKEKI, and EIAALEKEIAALEKEIAALEKEI, respectively. For the SHD fusions, Kar3 was fused to the basic helix, and Vik1 to the acidic helix.