title: Determination of peptide backbone torsion angles using double-quantum dipolar recoupling solid-state NMR spectroscopy.
abstract: Several approaches for utilizing dipolar recoupling solid-state NMR (ssNMR) techniques to determine local structure at high resolution in peptides and proteins have been developed. However, many of these techniques measure only one torsion angle or are accurate for only certain classes of secondary structure. Additionally, the efficiency with which these dipolar recoupling experiments suppress the deleterious effects of chemical shift anisotropy (CSA) at high magnetic field strengths varies. Dipolar recoupling with a windowless sequence (DRAWS) has proven to be an effective pulse sequence for exciting double-quantum (DQ) coherences between adjacent carbonyl carbons along the peptide backbone. By allowing this DQ coherence to evolve, it is possible to measure the relative orientations of the CSA tensors and subsequently use this information to determine the Ramachandran torsion angles phi and psi. Here, we explore the accuracies of the assumptions made in interpreting DQ-DRAWS data and demonstrate their fidelity in measuring torsion angles corresponding to a variety of secondary structures irrespective of hydrogen-bonding patterns. It is shown how a simple choice of isotopic labels and experimental conditions allows accurate measurement of backbone secondary structures without any prior knowledge. This approach is considerably more sensitive for determining structure in helices and has comparable accuracy for beta-sheet and extended conformations relative to other methods. We also illustrate the ability of DQ-DRAWS to distinguish between structures in heterogeneous samples.
authors.affiliation: Department of Chemistry and Biochemistry, 119 Woodland Street, Oberlin College, Oberlin, Ohio 44074, USA. manish.mehta@oberlin.edu
authors.forename: Manish A
authors.initials: MA
authors.lastname: Mehta
authors.forename: Matthew T
authors.initials: MT
authors.lastname: Eddy
authors.forename: Seth A
authors.initials: SA
authors.lastname: McNeill
authors.forename: Frank D
authors.initials: FD
authors.lastname: Mills
authors.forename: Joanna R
authors.initials: JR
authors.lastname: Long
mesh_terms: D003198:Computer Simulation; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D009842:Oligopeptides / Q000737:chemistry*; D017433:Protein Structure, Secondary; D011789:Quantum Theory; D014961:X-Ray Diffraction
chemical_list: D009842:Oligopeptides