Supplementary Figure 2: Flow chart of iterative EC-NMR analysis process.
From: Protein structure determination by combining sparse NMR data with evolutionary couplings
The EC-NMR analysis process has been fully automated by integration into the automated NOESY NMR data assignment program ASDP16,17. NOESY cross peak and NMR resonance assignments for 1H-15N and (where available) 1H-13C methyl resonances are combined with high-scoring EC contacts predicted from sequence co-variance analysis9,10. Specific restraints are identified by ASDP from these data, and used to generate structural models with the program CYANA18. Initial structures are then used to assess false-positive EC contacts and false-NOESY peaks (i.e. noise peaks), and to identify consistent Residue Pair Contacts (RPCs). Once the analysis converges, specific interatomic distance restraints are generated from the final RPC list and NOESY cross peak assignments, and used for restrained minimization of the EC-NMR structures with the program Rosetta19.
