title: Solid-state NMR structure determination from diagonal-compensated, sparsely nonuniform-sampled 4D proton-proton restraints.
abstract: We report acquisition of diagonal-compensated protein structural restraints from four-dimensional solid-state NMR spectra on extensively deuterated and (1)H back-exchanged proteins. To achieve this, we use homonuclear (1)H-(1)H correlations with diagonal suppression and nonuniform sampling (NUS). Suppression of the diagonal allows the accurate identification of cross-peaks which are otherwise obscured by the strong autocorrelation or whose intensity is biased due to partial overlap with the diagonal. The approach results in unambiguous spectral interpretation and relatively few but reliable restraints for structure calculation. In addition, the diagonal suppression produces a spectrum with low dynamic range for which ultrasparse NUS data sets can be readily reconstructed, allowing straightforward application of NUS with only 2% sampling density with the advantage of more heavily sampling time-domain regions of high signal intensity. The method is demonstrated here for two proteins, α-spectrin SH3 microcrystals and hydrophobin functional amyloids. For the case of SH3, suppression of the diagonal results in facilitated identification of unambiguous restraints and improvement of the quality of the calculated structural ensemble compared to nondiagonal-suppressed 4D spectra. For the only partly assigned hydrophobin rodlets, the structure is yet unknown. Applied to this protein of biological significance with large inhomogeneous broadening, the method allows identification of unambiguous crosspeaks that are otherwise obscured by the diagonal.
authors.affiliation: Max-Planck Institute for Biophysical Chemistry , Am Fassberg 11, 37077 Göttingen, Germany.
authors.forename: Rasmus
authors.initials: R
authors.lastname: Linser
authors.forename: Benjamin
authors.initials: B
authors.lastname: Bardiaux
authors.forename: Loren B
authors.initials: LB
authors.lastname: Andreas
authors.forename: Sven G
authors.initials: SG
authors.lastname: Hyberts
authors.forename: Vanessa K
authors.initials: VK
authors.lastname: Morris
authors.forename: Guido
authors.initials: G
authors.lastname: Pintacuda
authors.forename: Margaret
authors.initials: M
authors.lastname: Sunde
authors.forename: Ann H
authors.initials: AH
authors.lastname: Kwan
authors.forename: Gerhard
authors.initials: G
authors.lastname: Wagner
mesh_terms: D000682:Amyloid / Q000737:chemistry; D016477:Artifacts; D009682:Magnetic Resonance Spectroscopy; D008958:Models, Molecular; D011522:Protons; D013049:Spectrin / Q000737:chemistry; D018909:src Homology Domains
chemical_list: D000682:Amyloid; D011522:Protons; D013049:Spectrin