Validation of protein backbone structures calculated from NMR angular restraints using Rosetta

NMR “Crystallography” for Uniformly (13C, 15N)‐Labeled Oriented Membrane Proteins

“…rithm. [37] The details of the structural fitting procedure are provided in Section S2.2 of the Supporting Information. Briefly explained, the algorithm iteratively finds the values for the two pairs of torsion angles F and Y that are consistent with the sets of NMR angular restraints measured for three adjacent peptide planes (in this case 15 N CSA, 1 H-15 N, and 1 H a -13 C a dipolar couplings).…”

NMR “Crystallography” for Uniformly (13C, 15N)‐Labeled Oriented Membrane Proteins

“…Motions of the more dynamic (while structured) domains can potentially be accounted for in a manner similar to the residual dipolar couplings, that is, by introducing the additional local axial and rhombic components, which lies beyond the scope of the present paper. Ten thousand (10 000) structural solutions satisfying the three experimental angular restraints have been calculated using our recently developed structure‐fitting algorithm . The details of the structural fitting procedure are provided in Section S2.2 of the Supporting Information.…”

“…Ten thousand (10 000) structural solutions satisfying the three experimental angular restraints have been calculated using our recently developed structure-fitting algo- Angewandte Chemie Zuschriften rithm. [37] The details of the structural fitting procedure are provided in Section S2.2 of the Supporting Information. Briefly explained, the algorithm iteratively finds the values for the two pairs of torsion angles F and Y that are consistent with the sets of NMR angular restraints measured for three adjacent peptide planes (in this case 15 N CSA, 1 H-15 N, and 1 H a -13 C a dipolar couplings).…”

“…Structure determination of membrane proteins by OS-NMR will likely require measurements of additional angular restraints, for which doubly ( 15 N, 13 C)labeled protein samples would need to be employed. [35] Structure calculations from fitting synthetic multidimensional OS-NMR data [36,37] have identified 1 H a -13 C a dipolar couplings as a critical third spectroscopic dimension, which would enable the elucidation of the backbone folds of MPs without assuming any secondary structure elements a priori. The chiral nature of the 1 H a -13 C a dipolar couplings in the predominantly l-amino acids largely removes the orientational degeneracies associated with the second-rank 1 H- 15 N dipolar and 15 N CSA interactions.…”