title: Proton-detected scalar coupling based assignment strategies in MAS solid-state NMR spectroscopy applied to perdeuterated proteins.
abstract: Assignment of proteins in MAS (magic angle spinning) solid-state NMR relies so far on correlations among heteronuclei. This strategy is based on well dispersed resonances in the (15)N dimension. In many complex cases like membrane proteins or amyloid fibrils, an additional frequency dimension is desirable in order to spread the amide resonances. We show here that proton detected HNCO, HNCA, and HNCACB type experiments can successfully be implemented in the solid-state. Coherences are sufficiently long lived to allow pulse schemes of a duration greater than 70 ms before incrementation of the first indirect dimension. The achieved resolution is comparable to the resolution obtained in solution-state NMR experiments. We demonstrate the experiments using a triply labeled sample of the SH3 domain of chicken alpha-spectrin, which was re-crystallized in H(2)O/D(2)O using a ratio of 1/9. We employ paramagnetic relaxation enhancement (PRE) using EDTA chelated Cu(II) to enable rapid data acquisition.
authors.affiliation: Leibniz-Institut für Molekulare Pharmakologie (FMP), Campus Berlin-Buch, Robert-Rössle Strasse 10, 13125 Berlin, Germany.
authors.forename: Rasmus
authors.initials: R
authors.lastname: Linser
authors.forename: Uwe
authors.initials: U
authors.lastname: Fink
authors.forename: Bernd
authors.initials: B
authors.lastname: Reif
mesh_terms: D000818:Animals; D002247:Carbon Isotopes; D002645:Chickens; D003300:Copper; D003903:Deuterium / Q000737:chemistry; D004492:Edetic Acid; D009587:Nitrogen Isotopes; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D011487:Protein Conformation; D011522:Protons; D013049:Spectrin / Q000737:chemistry*
chemical_list: D002247:Carbon Isotopes; D009587:Nitrogen Isotopes; D011522:Protons; D013049:Spectrin; D003300:Copper; D004492:Edetic Acid; D003903:Deuterium