title: 4D APSY-HBCB(CG)CDHD experiment for automated assignment of aromatic amino acid side chains in proteins.
abstract: A four-dimensional (4D) APSY (automated projection spectroscopy)-HBCB(CG)CDHD experiment is presented. This 4D experiment correlates aromatic with aliphatic carbon and proton resonances from the same amino acid side chain of proteins in aqueous solution. It thus allows unambiguous sequence-specific assignment of aromatic amino acid ring signals based on backbone assignments. Compared to conventional 2D approaches, the inclusion of evolution periods on (1)H(β) and (13)C(δ) efficiently removes overlaps, and provides two additional frequencies for consequent automated or manual matching. The experiment was successfully applied to three proteins with molecular weights from 6 to 13 kDa. For the complementation of the assignment of the aromatic resonances, TOCSY- or COSY-based versions of a 4D APSY-HCCH(aro) sequence are proposed.
authors.affiliation: Institute of Molecular Biology and Biophysics, ETH Zurich, 8093 Zurich, Switzerland.
authors.forename: Barbara
authors.initials: B
authors.lastname: Krähenbühl
authors.forename: Sebastian
authors.initials: S
authors.lastname: Hiller
authors.forename: Gerhard
authors.initials: G
authors.lastname: Wider
mesh_terms: D024322:Amino Acids, Aromatic / Q000737:chemistry*; D002247:Carbon Isotopes; D008970:Molecular Weight; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D011487:Protein Conformation; D011506:Proteins / Q000737:chemistry*; D011522:Protons
chemical_list: D024322:Amino Acids, Aromatic; D002247:Carbon Isotopes; D011506:Proteins; D011522:Protons