title: 5D 13C-detected experiments for backbone assignment of unstructured proteins with a very low signal dispersion.
abstract: Two novel 5D NMR experiments (CACONCACO, NCOCANCO) for backbone assignment of disordered proteins are presented. The pulse sequences exploit relaxation properties of the unstructured proteins and combine the advantages of (13)C-direct detection, non-uniform sampling, and longitudinal relaxation optimization to maximize the achievable resolution and minimize the experimental time. The pulse sequences were successfully tested on the sample of partially disordered delta subunit from RNA polymerase from Bacillus subtilis. The unstructured part of this 20 kDa protein consists of 81 amino acids with frequent sequential repeats. A collection of 0.0003% of the data needed for a conventional experiment with linear sampling was sufficient to perform an unambiguous assignment of the disordered part of the protein from a single 5D spectrum.
authors.affiliation: Faculty of Science, NCBR, and CEITEC, Masaryk University, Brno, Czech Republic.
authors.forename: Jiří
authors.initials: J
authors.lastname: Nováček
authors.forename: Anna
authors.initials: A
authors.lastname: Zawadzka-Kazimierczuk
authors.forename: Veronika
authors.initials: V
authors.lastname: Papoušková
authors.forename: Lukáš
authors.initials: L
authors.lastname: Zídek
authors.forename: Hana
authors.initials: H
authors.lastname: Sanderová
authors.forename: Libor
authors.initials: L
authors.lastname: Krásný
authors.forename: Wiktor
authors.initials: W
authors.lastname: Koźmiński
authors.forename: Vladimír
authors.initials: V
authors.lastname: Sklenář
mesh_terms: D000595:Amino Acid Sequence; D001412:Bacillus subtilis / Q000201:enzymology; D001426:Bacterial Proteins / Q000737:chemistry*; D002247:Carbon Isotopes; D012321:DNA-Directed RNA Polymerases / Q000737:chemistry*; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D011487:Protein Conformation; D011506:Proteins / Q000737:chemistry*
chemical_list: D001426:Bacterial Proteins; D002247:Carbon Isotopes; D011506:Proteins; D012321:DNA-Directed RNA Polymerases