A dipolar coupling based strategy for simultaneous resonance assignment and structure determination of protein backbones

A new approach for simultaneous protein backbone resonance assignment and s tructure determination by NMR is introduced. This approach relies on recent advances in high-resolution NMR spectroscopy that allow observation of ani sotropic interactions, such as dipolar couplings. from proteins partially a ligned in field ordered media. Residual dipolar couplings are used for both geometric information and a filter in the assembly of residues in a sequen tial manner. Experimental data were collected in less than one week on a sm all redox protein, rubredoxin, that was N-15 enriched but not enriched abov e 1% natural abundance in C-13. Given the acceleration possible with partia l 13C enrichment, the protocol described should provide a very rapid route to protein structure determination. This is critical for the structural gen omics initiative where protein expression and structural determination in a high-throughput manner will be needed.