Crystal structure of a sweet tasting protein thaumatin I, at 1·65 Å resolution

Abstract

The crystal structure of thaumatin I, a potently sweet protein isolated from the fruits of the West African shrub, Thaumatococcus danielli Benth, has been refined at a resolution better than 1·65 Å using a combination of energy minimization and stereochemically restrained least-squares methods. The final model consists of all 207 amino acids, 28 alternate amino acid conformers and 236 waters, with a crystallographic R-factor of 0·145 for 19,877 reflections having F > 4σ F between 10·0 Å and 1·65 Å ( R = 0.167 for all 24,022 reflections). The model has good stereochemistry, with root-mean-square deviations from ideal values for bond and angle distances of 0·014 Å and 0·029 Å, respectively. 74The estimated root-mean-square co-ordinate error is 0·15 Å. The current model confirms the previously reported 3·1 Å C α trace in both main chain connectivity and disulfide topology, including two disulfide bonds, that differed from the earlier reported biochemical determination. The structure contains three domains. The core of the molecule consists of an eleven-stranded, flattened β-sandwich folded into two Greek key motifs. All β-strands in this sandwich are antiparallel except the parallel N-terminal and the C-terminal strands. The average hydrogen bond length in this sandwich is 2·89 Å, with an angle of 155·1 °. Two β-bulges are found in one of the sheets. The second domain consists of two β-strands forming a β-ribbon and connected by an ω-loop, and contains a proline residue in cis conformation. This structural motif folds back against the main sandwich to form a smaller sandwich-like structure. The third domain is a disulfide-rich region stretching away from the sandwich portion of the molecule. It contains one α-helix and three short helical fragments. Two of the helical segments are connected by an unusually sharp turn, stabilized by a disulfide bridge. One of the three disulfide bonds in this domain takes on two conformations.