An efficient high-throughput protocol based on 2D-HN(C)N for unambiguous H-N and N-15 backbone assignment in small folded proteins in less than a day

作者：Kumar Dinesh; Hosur Ramakrishna V^*

来源：Current Science, 2010, 99(11): 1581-1586.

• Backbone assignment
• check points
• sequential amide correlation
• structural proteomics

An efficient high-throughput method for sequential assignment of backbone H-1(N) and N-15, atoms in less than a day (using 2-4 2D spectra; total data collection in 4-8 h) has been proposed here. This is based on sequential correlations and specific patterns of peaks around the glycines, alanines, serines/threonines (internal check points) observable in the F-1-F-3 projection planes of the 3D-HN(C)N spectral variants. The F-2-F-3 projection planes of the spectra provide unique identification of the check points. The protocol has been demonstrated on two C-13/N-15-labelled proteins: ubiquitin and calbindin-D9k. In either case complete H-N and N-15 backbone assignments were obtained in less than a day each. The method would be valuable for NMR structural studies of small, well-folded proteins.

• 出版日期2010-12-10