Journal of Biomolecular NMR

, Volume 60, Issue 4, pp 209–218

“CON-CON” assignment strategy for highly flexible intrinsically disordered proteins

  • Alessandro Piai
  • Tomáš Hošek
  • Leonardo Gonnelli
  • Anna Zawadzka-Kazimierczuk
  • Wiktor Koźmiński
  • Bernhard Brutscher
  • Wolfgang Bermel
  • Roberta Pierattelli
  • Isabella C. Felli
Article

DOI: 10.1007/s10858-014-9867-6

Cite this article as:
Piai, A., Hošek, T., Gonnelli, L. et al. J Biomol NMR (2014) 60: 209. doi:10.1007/s10858-014-9867-6

Abstract

Intrinsically disordered proteins (IDPs) are a class of highly flexible proteins whose characterization by NMR spectroscopy is complicated by severe spectral overlaps. The development of experiments designed to facilitate the sequence-specific assignment procedure is thus very important to improve the tools for the characterization of IDPs and thus to be able to focus on IDPs of increasing size and complexity. Here, we present and describe the implementation of a set of novel 1H-detected 5D experiments, (HACA)CON(CACO)NCO(CA)HA, BT-(H)NCO(CAN)CONNH and BT-HN(COCAN)CONNH, optimized for the study of highly flexible IDPs that exploit the best resolved correlations, those involving the carbonyl and nitrogen nuclei of neighboring amino acids, to achieve sequence-specific resonance assignment. Together with the analogous recently proposed pulse schemes based on 13C detection, they form a complete set of experiments for sequence-specific assignment of highly flexible IDPs. Depending on the particular sample conditions (concentration, lifetime, pH, temperature, etc.), these experiments present certain advantages and disadvantages that will be discussed. Needless to say, that the availability of a variety of complementary experiments will be important for accurate determination of resonance frequencies in complex IDPs.

Keywords

Intrinsically disordered proteins 13C detection 1HN detection 1Hα detection NUS Multidimensional NMR experiment BEST-TROSY Backbone assignment 

Supplementary material

10858_2014_9867_MOESM1_ESM.pdf (708 kb)
Supplementary material 1 (PDF 708 kb)

Copyright information

© Springer Science+Business Media Dordrecht 2014

Authors and Affiliations

  • Alessandro Piai
    • 1
  • Tomáš Hošek
    • 1
  • Leonardo Gonnelli
    • 1
  • Anna Zawadzka-Kazimierczuk
    • 2
  • Wiktor Koźmiński
    • 2
  • Bernhard Brutscher
    • 3
  • Wolfgang Bermel
    • 4
  • Roberta Pierattelli
    • 1
  • Isabella C. Felli
    • 1
  1. 1.CERM and Department of ChemistryUniversity of FlorenceSesto FiorentinoItaly
  2. 2.Faculty of Chemistry, Biological and Chemical Research CentreUniversity of WarsawWarsawPoland
  3. 3.Institut de Biologie Structurale, Université Grenoble 1CNRSGrenoble Cedex 1France
  4. 4.Bruker BioSpin GmbHRheinstettenGermany

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