Journal of Biomolecular NMR

, Volume 50, Issue 1, pp 1–11 | Cite as

5D 13C-detected experiments for backbone assignment of unstructured proteins with a very low signal dispersion

  • Jiří Nováček
  • Anna Zawadzka-Kazimierczuk
  • Veronika Papoušková
  • Lukáš Žídek
  • Hana Šanderová
  • Libor Krásný
  • Wiktor Koźmiński
  • Vladimír Sklenář
Communication

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 13C-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.

Keywords

Intrinsically disordered proteins Non-uniform sampling 13C detection Longitudinal relaxation optimization Backbone assignment 

Supplementary material

10858_2011_9496_MOESM1_ESM.pdf (313 kb)
PDF (312 KB)

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Copyright information

© Springer Science+Business Media B.V. 2011

Authors and Affiliations

  • Jiří Nováček
    • 1
  • Anna Zawadzka-Kazimierczuk
    • 2
  • Veronika Papoušková
    • 1
  • Lukáš Žídek
    • 1
  • Hana Šanderová
    • 3
  • Libor Krásný
    • 3
  • Wiktor Koźmiński
    • 2
  • Vladimír Sklenář
    • 1
  1. 1.Faculty of Science, NCBR, and CEITECMasaryk UniversityBrnoCzech Republic
  2. 2.Faculty of ChemistryUniversity of WarsawWarsawPoland
  3. 3.Laboratory of Molecular Genetics of Bacteria and Department of BacteriologyInstitute of Microbiology, Academy of Sciences of the Czech RepublicPragueCzech Republic

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