Abstract An experiment is presented to determine 3JHNHα coupling constants, with significant advantages for applications to unfolded proteins. The determination of coupling constants for the peptide chain using 1D 1H, or 2D and 3D 1H-15N correlation spectroscopy is often hampered by extensive resonance overlap when dealing with flexible, disordered proteins. In the experiment detailed here, the overlap problem is largely circumvented by recording 1H-13C� correlation spectra, which demonstrate superior resolution for unfolded proteins. J-coupling constants are extracted from the peak intensities in a pair of 2D spin-echo difference experiments, affording rapid acquisition of the coupling data. In an application to the cytoplasmic domain of human neuroligin-3 (hNlg3cyt) data were obtained for 78 residues, compared to 54 coupling constants obtained from a 3D HNHA experiment. The coupling constants suggest that hNlg3cyt is intrinsically disordered, with little propensity for structure.
Content Type Journal Article
Pages 343-349
DOI 10.1007/s10858-009-9382-3
Authors
Renee Otten, University of Groningen Department of Biophysical Chemistry, Groningen Biomolecular Sciences and Biotechnology Institute Nijenborgh 4 9747 AG Groningen The Netherlands
Kathleen Wood, University of Groningen Department of Biophysical Chemistry, Groningen Biomolecular Sciences and Biotechnology Institute Nijenborgh 4 9747 AG Groningen The Netherlands
Frans A. A. Mulder, University of Groningen Department of Biophysical Chemistry, Groningen Biomolecular Sciences and Biotechnology Institute Nijenborgh 4 9747 AG Groningen The Netherlands
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