Skip to main content
SpringerLink
Log in

1H, 13C and 15N resonance assignment of a 114-residue fragment of Engrailed 2 homeoprotein, a partially disordered protein

  • Article
  • Published:
Biomolecular NMR Assignments Aims and scope Submit manuscript

Abstract

Engrailed 2 is a transcription factor belonging to the class of homeoproteins. These proteins possess a 60-residue DNA binding globular domain and play an important role in the early stages of development. We expressed and purified a 13.4 kDa fragment of Engrailed 2, which comprises a 54-residue N-terminal extension in addition to the homeodomain region. Almost all backbone and side-chain resonances have been assigned. The weak dispersion in the proton dimension of the 1H–15N HSQC spectrum indicates the presence of disordered regions that do not belong to the homeodomain. This work is a first step toward the NMR investigation of the structure and dynamics of Engrailed 2 protein that contains a well-structured globular domain and partially disordered regions.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1

Similar content being viewed by others

References

  • Alberi L, Sgado P, Simon HH (2004) Engrailed genes are cell-autonomously required to prevent apoptosis in mesencephalic dopaminergic neurons. Development 131:3229–3236

    Article  Google Scholar 

  • Delaglio F, Grzesiek S, Vuister GW, Zhu G, Pfeifer J, Bax A (1995) NMRPipe: a multidimensional spectral processing system based on UNIX pipes. J Biomol NMR 6:277–293

    Article  Google Scholar 

  • Foucher I, Montesinos ML, Volovitch M, Prochiantz A, Trembleau A (2003) Joint regulation of the MAP1B promoter by HNF3beta/Foxa2 and Engrailed is the result of a highly conserved mechanism for direct interaction of homeoproteins and Fox transcription factors. Development 130:1867–1876

    Article  Google Scholar 

  • Fraenkel E, Rould MA, Chambers KA, Pabo CO (1998) Engrailed homeodomain-DNA complex at 2.2 Å resolution: a detailed view of the interface and comparison with other engrailed structures. J Mol Biol 284:351–361

    Article  Google Scholar 

  • Gehring WJ (1987) Homeo boxes in the study of development. Science 236:1245–1252

    Article  ADS  Google Scholar 

  • Gehring WJ, Affolter M, Buerglin T (1994) Homeodomain proteins. Ann Rev Biochem 63:487–526

    Article  Google Scholar 

  • Joyner AL (1996) Engrailed, Wnt and Pax genes regulate midbrain-hindbrain development. Trends Genet 12:15–20

    Article  Google Scholar 

  • Kazimierczuk K, Zawadzka A, Kozminski W (2009) Narrow peaks and high dimensionalities: exploiting the advantages of random sampling. J Magn Reson 197:210–228

    Article  ADS  Google Scholar 

  • Maizel A, Tassetto M, Filhol O, Cochet C, Prochiantz A, Joliot A (2002) Engrailed homeoprotein secretion is a regulated process. Development 129:3545–3553

    Google Scholar 

  • Marley J, Lu M, Bracken C (2001) A method for efficient isotopic labelling of recombinant proteins. J Biomol NMR 20:71–75

    Article  Google Scholar 

  • McGinnis W, Levine MS, Hafen E, Kuroiwa A, Gehring WJ (1984) A conserved DNA sequence in homeotic genes of the Drosophila Antennapedia and bithorax complex. Nature 308:428–433

    Article  ADS  Google Scholar 

  • Piper DE, Batchelor AH, Chang CP, Cleary ML, Wolberger C (1999) Structure of the HoxB1-Pbx1 heterodimer bound to DNA: role of the hexapeptide and the fourth homeodomain helix in complex formation. Cell 96:587–597

    Article  Google Scholar 

  • Sattler M, Schleucher J, Griesinger C (1999) Heteronuclear multidimensional NMR experiments for the structure determination of proteins in solution employing pulsed field gradients. Prog Nucl Magn Reson Spectrosc 34:93–158

    Article  Google Scholar 

  • Simon HH, Saueressig H, Wurst W, Goulding MD, O’Leary DD (2001) Fate of midbrain dopaminergic neurons controlled by the engrailed genes. J Neurosci 21:3126–3134

    Google Scholar 

  • Sonnier L, Le Pen G, Hartmann A, Bizot JC, Trovero F, Krebs MO, Prochiantz A (2007) Progressive loss of dopaminergic neurons in the ventral midbrain of adult mice heterozygote for Engrailed1. J Neurosci 27:1063–1071

    Article  Google Scholar 

  • Vranken WF, Boucher W, Stevens TJ et al (2005) The CCPN data model for NMR spectroscopy: development of a software pipeline. Proteins 59:687–696

    Article  Google Scholar 

Download references

Acknowledgments

Financial support by the Access to Research Infrastructures activity of the 7th Framework Programme of the EU (Contract 228461, EAST-NMR) for conducting research in Warsaw is gratefully acknowledged. Wiktor Kozminski and Jan Stanek (Department of Chemistry, University of Warsaw) are deeply acknowledged for their help in NMR data collection and fruitful discussions. We thank Alain Prochiantz (UMR 7233 CNRS, Collège de France) for kindly providing the expression plasmid.

Author information

Authors and Affiliations

  1. UPMC Univ Paris 06, 4 Place Jussieu, 75252, Paris, Cedex 05, France

    Rafal Augustyniak, Fabien Ferrage, Geoffrey Bodenhausen & Olivier Lequin

  2. Département de chimie, Ecole Normale Supérieure, 24 rue Lhomond, 75231, Paris Cedex 05, France

    Rafal Augustyniak, Fabien Ferrage, Geoffrey Bodenhausen & Olivier Lequin

  3. UMR 7203 CNRS-UPMC-ENS, Paris Cedex 05, France

    Rafal Augustyniak, Fabien Ferrage, Geoffrey Bodenhausen & Olivier Lequin

  4. Groupe de RMN Biomédicale, Laboratoire de Synthèse et Physico-Chimie de Molécules d’Intérêt Biologique, UMR CNRS 5068 CNRS, Université de Toulouse, 118 route de Narbonne, 31062, Toulouse Cedex 09, France

    Stéphane Balayssac

  5. Institut des Sciences et Ingénierie Chimiques, BCH, Ecole Polytechnique Fédérale de Lausanne, 1015, Lausanne, Switzerland

    Geoffrey Bodenhausen

Authors
  1. Rafal Augustyniak
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Stéphane Balayssac
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Fabien Ferrage
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Geoffrey Bodenhausen
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Olivier Lequin
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Olivier Lequin.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Augustyniak, R., Balayssac, S., Ferrage, F. et al. 1H, 13C and 15N resonance assignment of a 114-residue fragment of Engrailed 2 homeoprotein, a partially disordered protein. Biomol NMR Assign 5, 229–231 (2011). https://doi.org/10.1007/s12104-011-9306-5

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12104-011-9306-5

Keywords

Search

Navigation