Patent ID: 7365539

Claim:
A method for sequentially assigning chemical shift values of an α-proton, 1 H α , an α-carbon, 13 C α , a polypeptide backbone carbonyl carbon, 13 C′, a polypeptide backbone amide nitrogen, 15 N, and a polypeptide backbone amide proton, 1 H N , of a protein molecule comprising: providing a protein sample; conducting a set of G matrix Fourier transformation (GFT) nuclear magnetic resonance (NMR) experiments on the protein sample comprising: (1) a (5,2)D [ HACACON HN] GFT NMR experiment to measure and connect the chemical shift values of the α-proton of amino acid residue i−1, 1 H αi−1 , the α-carbon of amino acid residue i−1, 13 C α i−1 , the polypeptide backbone carbonyl carbon of amino acid residue i−1, 13 C′ i−1 , the polypeptide backbone amide nitrogen of amino acid residue i, 15 N i , and the polypeptide backbone amide proton of amino acid residue i, 1 H N i and (2) a (5,2)D [ HACA , CON HN] GFT NMR experiment to measure and connect the chemical shift values of 1 H α i−1 , 13 C α i−1 , 13 C′ i−1 , the polypeptide backbone amide nitrogen of amino acid residue i−1, 15 N i−1 , and the polypeptide backbone amide proton of amino acid residue i−1, 1 H N i−1 ; and obtaining sequential assignments of the chemical shift values of 1 H α , 13 C α , 13 C′, 15 N, and 1 H N by (i) matching the chemical shift values of 1 H α i−1 , 13 C α i−1 , and 13 C′ i−1 measured by said (5,2)D [ HACACON HN] GFT NMR experiment with the chemical shift values of 1 H α i−1 , 13 C α i−1 , and 13 C′ i−1 measured by said (5,2)D [ HACA , CON HN] GFT NMR experiment, (ii) using the chemical shift values of 1 H α i−1 , 13 C α i−1 , and 13 C′ i−1 to identify the type of amino acid residue i−1, and (iii) mapping sets of sequentially connected chemical shift values to the amino acid sequence of the polypeptide chain.