HET-SOFAST NMR for fast detection of structural compactness and heterogeneity along polypeptide chains.
Structure elucidation of proteins by either NMR or X-ray crystallography often requires the screening of a large number of samples for promising protein constructs and optimum solution conditions. For large-scale screening of protein samples in solution, robust methods are needed that allow a rapid assessment of the folding of a polypeptide under diverse sample conditions. Here we present HET-SOFAST NMR, a highly sensitive new method for semi-quantitative characterization of the structural compactness and heterogeneity of polypeptide chains in solution. On the basis of one-dimensional 1H HET-SOFAST NMR data, obtained on well-folded, molten globular, partially- and completely unfolded proteins, we define empirical thresholds that can be used as quantitative benchmarks for protein compactness. For 15N-enriched protein samples, two-dimensional 1H-15N HET-SOFAST correlation spectra provide site-specific information about the structural heterogeneity along the polypeptide chain.
Schanda
Paul
P
Institut de Biologie Structurale, Jean-Pierre Ebel C.N.R.S.-C.E.A.-UJF, 41, rue Jules Horowitz, 38027 Grenoble Cedex 1, France.
Forge
Vincent
V
Brutscher
Bernhard
B
D006801:Humans; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D010455:Peptides / Q000737:chemistry*; D011487:Protein Conformation; D017510:Protein Folding; D012680:Sensitivity and Specificity; D012996:Solutions / Q000737:chemistry
D010455:Peptides; D012996:Solutions