title: Structural constraints for the Crh protein from solid-state NMR experiments.
abstract: We demonstrate that short, medium and long-range constraints can be extracted from proton mediated, rare-spin detected correlation solid-state NMR experiments for the microcrystalline 10.4 x 2 kDa dimeric model protein Crh. Magnetization build-up curves from cross signals in NHHC and CHHC spectra deliver detailed information on side chain conformers and secondary structure for interactions between spin pairs. A large number of medium and long-range correlations can be observed in the spectra, and an analysis of the resolved signals reveals that the constraints cover the entire sequence, also including inter-monomer contacts between the two molecules forming the domain-swapped Crh dimer. Dynamic behavior is shown to have an impact on cross signals intensities, as indicated for mobile residues or regions by contacts predicted from the crystal structure, but absent in the spectra. Our work validates strategies involving proton distance measurements for large and complex proteins as the Crh dimer, and confirms the magnetization transfer properties previously described for small molecules in solid protein samples.
authors.affiliation: Institut de Biologie et Chimie des Protéines, UMR 5086 C.N.R.S./Université de Lyon, Lyon Cedex 07, France.
authors.forename: Carole
authors.initials: C
authors.lastname: Gardiennet
authors.forename: Antoine
authors.initials: A
authors.lastname: Loquet
authors.forename: Manuel
authors.initials: M
authors.lastname: Etzkorn
authors.forename: Henrike
authors.initials: H
authors.lastname: Heise
authors.forename: Marc
authors.initials: M
authors.lastname: Baldus
authors.forename: Anja
authors.initials: A
authors.lastname: Böckmann
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry*; D019281:Dimerization; D008958:Models, Molecular; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D010750:Phosphoproteins / Q000737:chemistry*; D011487:Protein Conformation; D011522:Protons
chemical_list: D001426:Bacterial Proteins; C118042:Crh protein, Bacillus subtilis; D010750:Phosphoproteins; D011522:Protons