Highly efficient NMR assignment of intrinsically disordered proteins: application to B- and T cell receptor domains

Linnéa Isaksson; Maxim Mayzel; Maria Saline; Anders Pedersen; Joakim Rosenlöw; Bernhard Brutscher; B Göran Karlsson; Vladislav Y Orekhov

doi:10.1371/journal.pone.0062947

Highly efficient NMR assignment of intrinsically disordered proteins: application to B- and T cell receptor domains

PLoS One. 2013 May 7;8(5):e62947. doi: 10.1371/journal.pone.0062947. Print 2013.

Authors

Linnéa Isaksson¹, Maxim Mayzel, Maria Saline, Anders Pedersen, Joakim Rosenlöw, Bernhard Brutscher, B Göran Karlsson, Vladislav Y Orekhov

Affiliation

¹ Swedish NMR Centre, University of Gothenburg, Gothenburg, Sweden.

Abstract

We present an integrated approach for efficient characterization of intrinsically disordered proteins. Batch cell-free expression, fast data acquisition, automated analysis, and statistical validation with data resampling have been combined for achieving cost-effective protein expression, and rapid automated backbone assignment. The new methodology is applied for characterization of five cytosolic domains from T- and B-cell receptors in solution.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Amino Acid Motifs
Cytosol / metabolism
Humans
Intracellular Space / metabolism
Intrinsically Disordered Proteins / chemistry*
Intrinsically Disordered Proteins / metabolism
Ligands
Nuclear Magnetic Resonance, Biomolecular / methods*
Protein Structure, Tertiary
Receptors, Antigen, B-Cell / chemistry*
Receptors, Antigen, B-Cell / metabolism
Receptors, Antigen, T-Cell / chemistry*
Receptors, Antigen, T-Cell / metabolism
Signal Transduction

Substances

Intrinsically Disordered Proteins
Ligands
Receptors, Antigen, B-Cell
Receptors, Antigen, T-Cell

Grants and funding

The work was supported by the Swedish Research Council (621-2005-2951, 2011-5994), Wenner-Gren Foundation (Stockholm), and the European Commis-sion (FP7-I3-Bio-NMR project No. 261863 and FP7-ITN-IDPbyNMR contract No. 264257). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.