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

• Published in: PloS one Vol. 8; no. 5; p. e62947
• Main Authors: Isaksson, Linnéa, Mayzel, Maxim, Saline, Maria, Pedersen, Anders, Rosenlöw, Joakim, Brutscher, Bernhard, Karlsson, B Göran, Orekhov, Vladislav Y
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 07.05.2013; Public Library of Science (PLoS)
• Subjects: Amino Acid Motifs; Biochemistry, Molecular Biology; Biology; Cellular signal transduction; Chemical Sciences; Chemistry; Cytosol; Cytosol - metabolism; Data acquisition; Data processing; E coli; Experiments; Freedom of speech; Humans; Intracellular Space; Intracellular Space - metabolism; Intrinsically Disordered Proteins; Intrinsically Disordered Proteins - chemistry; Intrinsically Disordered Proteins - metabolism; Kemi; Kinases; Life Sciences; Ligands; Lymphocytes B; NMR; Nuclear magnetic resonance; Nuclear Magnetic Resonance, Biomolecular; Nuclear Magnetic Resonance, Biomolecular - methods; Peptides; Physics; Protein folding; Protein Structure, Tertiary; Protein synthesis; Proteins; Receptors; Receptors, Antigen, B-Cell; Receptors, Antigen, B-Cell - chemistry; Receptors, Antigen, B-Cell - metabolism; Receptors, Antigen, T-Cell; Receptors, Antigen, T-Cell - chemistry; Receptors, Antigen, T-Cell - metabolism; Resampling; Signal Transduction; Spectrum analysis; Statistical analysis; Structural Biology; T cell receptors; T cells; T-cell receptor
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0062947

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.