title: Proton-detected MAS NMR experiments based on dipolar transfers for backbone assignment of highly deuterated proteins.
abstract: Proton-detected solid-state NMR was applied to a highly deuterated insoluble, non-crystalline biological assembly, the Salmonella typhimurium type iii secretion system (T3SS) needle. Spectra of very high resolution and sensitivity were obtained at a low protonation level of 10-20% at exchangeable amide positions. We developed efficient experimental protocols for resonance assignment tailored for this system and the employed experimental conditions. Using exclusively dipolar-based interspin magnetization transfers, we recorded two sets of 3D spectra allowing for an almost complete backbone resonance assignment of the needle subunit PrgI. The additional information provided by the well-resolved proton dimension revealed the presence of two sets of resonances in the N-terminal helix of PrgI, while in previous studies employing (13)C detection only a single set of resonances was observed.
authors.affiliation: Max Planck Institute for Biophysical Chemistry, Department of NMR-based Structural Biology, Am Fassberg 11, 37077 Göttingen, Germany.
authors.forename: Veniamin
authors.initials: V
authors.lastname: Chevelkov
authors.affiliation: Max Planck Institute for Biophysical Chemistry, Department of NMR-based Structural Biology, Am Fassberg 11, 37077 Göttingen, Germany.
authors.forename: Birgit
authors.initials: B
authors.lastname: Habenstein
authors.affiliation: Max Planck Institute for Biophysical Chemistry, Department of NMR-based Structural Biology, Am Fassberg 11, 37077 Göttingen, Germany.
authors.forename: Antoine
authors.initials: A
authors.lastname: Loquet
authors.affiliation: Max Planck Institute for Biophysical Chemistry, Department of NMR-based Structural Biology, Am Fassberg 11, 37077 Göttingen, Germany.
authors.forename: Karin
authors.initials: K
authors.lastname: Giller
authors.affiliation: Max Planck Institute for Biophysical Chemistry, Department of NMR-based Structural Biology, Am Fassberg 11, 37077 Göttingen, Germany.
authors.forename: Stefan
authors.initials: S
authors.lastname: Becker
authors.affiliation: Max Planck Institute for Biophysical Chemistry, Department of NMR-based Structural Biology, Am Fassberg 11, 37077 Göttingen, Germany. Electronic address: adla@nmr.mpibpc.mpg.de.
authors.forename: Adam
authors.initials: A
authors.lastname: Lange
mesh_terms: D000465:Algorithms; D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry* / Q000648:ultrastructure*; D003198:Computer Simulation; D003903:Deuterium / Q000737:chemistry*; D041961:Deuterium Exchange Measurement / Q000379:methods; D008956:Models, Chemical; D008969:Molecular Sequence Data; D066244:Proton Magnetic Resonance Spectroscopy / Q000379:methods*; D012486:Salmonella typhimurium / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D003903:Deuterium
keywords: Deuteration; Magic-angle spinning solid-state NMR; PrgI; Protein resonance assignment; Proton detection; Type 3 secretion system