title: Protein residue linking in a single spectrum for magic-angle spinning NMR assignment.
abstract: Here we introduce a new pulse sequence for resonance assignment that halves the number of data sets required for sequential linking by directly correlating sequential amide resonances in a single diagonal-free spectrum. The method is demonstrated with both microcrystalline and sedimented deuterated proteins spinning at 60 and 111 kHz, and a fully protonated microcrystalline protein spinning at 111 kHz, with as little as 0.5 mg protein sample. We find that amide signals have a low chance of ambiguous linkage, which is further improved by linking in both forward and backward directions. The spectra obtained are amenable to automated resonance assignment using general-purpose software such as UNIO-MATCH.
authors.affiliation: Centre de RMN à Très Hauts Champs, Institut des Sciences Analytiques (CNRS, ENS Lyon, UCB Lyon 1), Université de Lyon, 69100, Villeurbanne, France.
authors.forename: Loren B
authors.initials: LB
authors.lastname: Andreas
authors.forename: Jan
authors.initials: J
authors.lastname: Stanek
authors.forename: Tanguy
authors.initials: T
authors.lastname: Le Marchand
authors.forename: Andrea
authors.initials: A
authors.lastname: Bertarello
authors.forename: Diane
authors.initials: D
authors.lastname: Cala-De Paepe
authors.forename: Daniela
authors.initials: D
authors.lastname: Lalli
authors.forename: Magdaléna
authors.initials: M
authors.lastname: Krejčíková
authors.forename: Camille
authors.initials: C
authors.lastname: Doyen
authors.forename: Carl
authors.initials: C
authors.lastname: Öster
authors.forename: Benno
authors.initials: B
authors.lastname: Knott
authors.forename: Sebastian
authors.initials: S
authors.lastname: Wegner
authors.forename: Frank
authors.initials: F
authors.lastname: Engelke
authors.forename: Isabella C
authors.initials: IC
authors.lastname: Felli
authors.forename: Roberta
authors.initials: R
authors.lastname: Pierattelli
authors.forename: Nicholas E
authors.initials: NE
authors.lastname: Dixon
authors.forename: Lyndon
authors.initials: L
authors.lastname: Emsley
authors.forename: Torsten
authors.initials: T
authors.lastname: Herrmann
authors.forename: Guido
authors.initials: G
authors.lastname: Pintacuda
mesh_terms: D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D011506:Proteins / Q000737:chemistry*; D011522:Protons
chemical_list: D011506:Proteins; D011522:Protons