title: 13C spin dilution for simplified and complete solid-state NMR resonance assignment of insoluble biological assemblies.
abstract: A strategy for simplified and complete resonance assignment of insoluble and noncrystalline proteins by solid-state NMR (ssNMR) spectroscopy is presented. Proteins produced with [1-(13)C]- or [2-(13)C]glucose are very sparsely labeled, and the resulting 2D ssNMR spectra exhibit smaller line widths (by a factor of ∼2 relative to uniformly labeled proteins) and contain a reduced number of cross-peaks. This allows for an accelerated and straightforward resonance assignment without the necessity of time-consuming 3D spectroscopy or sophisticated pulse sequences. The strategy aims at complete backbone and side-chain resonance assignments based on bidirectional sequential walks. The approach was successfully demonstrated with the de novo assignment of the Type Three Secretion System PrgI needle protein. Using a limited set of simple 2D experiments, we report a 97% complete resonance assignment of the backbone and side-chain (13)C atoms.
authors.affiliation: Department of NMR-Based Structural Biology, Max Planck Institute for Biophysical Chemistry, Am Fassberg 11, 37077 Göttingen, Germany.
authors.forename: Antoine
authors.initials: A
authors.lastname: Loquet
authors.forename: Guohua
authors.initials: G
authors.lastname: Lv
authors.forename: Karin
authors.initials: K
authors.lastname: Giller
authors.forename: Stefan
authors.initials: S
authors.lastname: Becker
authors.forename: Adam
authors.initials: A
authors.lastname: Lange
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry*; D002247:Carbon Isotopes; D009682:Magnetic Resonance Spectroscopy / Q000592:standards; D049329:Nanostructures / Q000737:chemistry; D010316:Particle Size; D012015:Reference Standards; D012486:Salmonella typhimurium / Q000737:chemistry; D012995:Solubility
chemical_list: D001426:Bacterial Proteins; D002247:Carbon Isotopes