Efficient sequential assignments in proteins with reduced dimensionality 3D HN(CA)NH.
We present reduced dimensionality (RD) 3D HN(CA)NH for efficient sequential assignment in proteins. The experiment correlates the (15)N and (1)H chemical shift of a residue ('i') with those of its immediate N-terminal (i - 1) and C-terminal (i + 1) neighbors and provides four-dimensional chemical shift correlations rapidly with high resolution. An assignment strategy is presented which combines the correlations observed in this experiment with amino acid type information obtained from 3D CBCA(CO)NH. By classifying the 20 amino acid types into seven distinct categories based on (13)C(β) chemical shifts, it is observed that a stretch of five sequentially connected residues is sufficient to map uniquely on to the polypeptide for sequence specific resonance assignments. This method is exemplified by application to three different systems: maltose binding protein (42 kDa), intrinsically disordered domain of insulin-like growth factor binding protein-2 and Ubiquitin. Fast data acquisition is demonstrated using longitudinal (1)H relaxation optimization. Overall, 3D HN(CA)NH is a powerful tool for high throughput resonance assignment, in particular for unfolded or intrinsically disordered polypeptides.
NMR Research Centre, Indian Institute of Science, Bangalore, 560012, India.
Kousik
K
Chandra
Garima
G
Jaipuria
Divya
D
Shet
Hanudatta S
HS
Atreya
D000596:Amino Acids / Q000737:chemistry; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D018969:Insulin-Like Growth Factor Binding Proteins / Q000737:chemistry; D058619:Maltose-Binding Proteins / Q000737:chemistry; D015233:Models, Statistical; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D011506:Proteins / Q000737:chemistry*; D011994:Recombinant Proteins / Q000737:chemistry; D025801:Ubiquitin / Q000737:chemistry
D000596:Amino Acids; D018969:Insulin-Like Growth Factor Binding Proteins; D058619:Maltose-Binding Proteins; D011506:Proteins; D011994:Recombinant Proteins; D025801:Ubiquitin