title: 3D NMR spectroscopy for resonance assignment and structure elucidation of proteins under MAS: novel pulse schemes and sensitivity considerations.
abstract: Two types of 3D MAS NMR experiments are introduced, which combine standard (NC,CC) transfer schemes with (1H,1H) mixing to simultaneously detect connectivities and structural constraints of uniformly 15N,13C-labeled proteins with high spectral resolution. The homonuclear CCHHC and CCC experiments are recorded with one double-quantum evolution dimension in order to avoid a cubic diagonal in the spectrum. Depending on the second transfer step, spin systems or proton-proton contacts can be determined with reduced spectral overlap. The heteronuclear NHHCC experiment encodes NH-HC proton-proton interactions, which are indicative for the backbone conformation of the protein. The third dimension facilitates the identification of the amino acid spin system. Experimental results on U-[15N,13C]valine and U-[15N,13C]ubiquitin demonstrate their usefulness for resonance assignments and for the determination of structural constraints. Furthermore, we give a detailed analysis of alternative multidimensional sampling schemes and their effect on sensitivity and resolution.
authors.affiliation: Department for NMR-based Structural Biology, Max-Planck-Institute for Biophysical Chemistry, Am Fassberg 11, D-37077 Göttingen, Germany. hehe@nmr.mpibpc.mpg.de
authors.forename: Henrike
authors.initials: H
authors.lastname: Heise
authors.forename: Karsten
authors.initials: K
authors.lastname: Seidel
authors.forename: Manuel
authors.initials: M
authors.lastname: Etzkorn
authors.forename: Stefan
authors.initials: S
authors.lastname: Becker
authors.forename: Marc
authors.initials: M
authors.lastname: Baldus
mesh_terms: D000596:Amino Acids / Q000737:chemistry; D002247:Carbon Isotopes; D005449:Fluorenes / Q000737:chemistry; D015394:Molecular Structure; D009587:Nitrogen Isotopes; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D011506:Proteins / Q000737:chemistry; D011522:Protons; D012680:Sensitivity and Specificity; D025801:Ubiquitin / Q000737:chemistry*; D014633:Valine / Q000737:chemistry*
chemical_list: D000596:Amino Acids; D002247:Carbon Isotopes; D005449:Fluorenes; C016456:N(alpha)-fluorenylmethyloxycarbonylamino acids; D009587:Nitrogen Isotopes; D011506:Proteins; D011522:Protons; D025801:Ubiquitin; D014633:Valine