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Journal of Biomolecular NMR

, Volume 53, Issue 4, pp 293–301 | Cite as

Speeding up sequence specific assignment of IDPs

  • Wolfgang Bermel
  • Ivano Bertini
  • Isabella C. Felli
  • Leonardo Gonnelli
  • Wiktor Koźmiński
  • Alessandro Piai
  • Roberta Pierattelli
  • Jan Stanek
Article

Abstract

The characterization of intrinsically disordered proteins (IDPs) by NMR spectroscopy is made difficult by the extensive spectral overlaps. To overcome the intrinsic low-resolution of the spectra the introduction of high-dimensionality experiments is essential. We present here a set of high-resolution experiments based on direct 13C-detection which proved useful in the assignment of α-synuclein, a paradigmatic IDP. In particular, we describe the implementation of 4D HCBCACON, HCCCON, HCBCANCO, 4/5D HNCACON and HNCANCO and 3/4D HCANCACO experiments, specifically tailored for spin system identification and backbone resonances sequential assignment. The use of non-uniform-sampling in the indirect dimension and of the H-flip approach to achieve longitudinal relaxation enhancement rendered the experiments very practical.

Keywords

Intrinsically disordered proteins 13C detection Non-uniform sampling Multidimensional NMR experiment Backbone assignment Spin system identification 

Notes

Acknowledgments

This work has been supported in part by the EC 7th Framework program BioNMR, contract 261863 and by the EC Marie Curie ITN program (IDPbyNMR, contract 264257).

Supplementary material

10858_2012_9639_MOESM1_ESM.doc (1.3 mb)
Supplementary material 1 (DOC 1292 kb)

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Copyright information

© Springer Science+Business Media B.V. 2012

Authors and Affiliations

  • Wolfgang Bermel
    • 1
  • Ivano Bertini
    • 2
    • 3
  • Isabella C. Felli
    • 2
    • 3
  • Leonardo Gonnelli
    • 2
  • Wiktor Koźmiński
    • 4
  • Alessandro Piai
    • 2
  • Roberta Pierattelli
    • 2
    • 3
  • Jan Stanek
    • 4
  1. 1.Bruker BioSpin GmbHRheinstettenGermany
  2. 2.CERM, University of FlorenceSesto FiorentinoItaly
  3. 3.Department of Chemistry “Ugo Schiff”University of FlorenceSesto FiorentinoItaly
  4. 4.Faculty of ChemistryUniversity of WarsawWarsawPoland

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