Binding mode of brazzein to the taste receptor based on crystal structure and docking simulation

Several natural substances including protein produce sweet taste. Brazzein, derived from the plant Pentadipladra brazzeana, is one of the sweet proteins that bind to the taste receptor with stronger sweetness than sugar. Mutations of this protein affect its flavour, yielding higher sweetness in D29K and lower sweetness in R43A. To elucidate its sweet mechanism in the taste receptor, we determined the structures of two variants, D29K and R43A, to a resolution of 1.5 Å and 1.3 Å, respectively. Structures of the brazzein exhibit two α-helix and three β-sheets connected by four disulfide bonds with a significantly altered electrostatic distribution on the surface. Using the high-resolution structure data and models of the taste receptors T1R2 and T1R3 in the AlphaFold Protein Structure Database, we performed a docking calculation on the receptors and report that brazzein is bound between the two cysteine rich domains (CRDs) of the heterodimer protein complex. Substitution to lysine in D29K resulted in an increased number of hydrogen bonds in the T1R2 receptor, while substitution to alanine in R43A ablated a polar interaction in the T1R3 receptor. The significantly altered interaction of the variants at the interface is consistent with a change of the sweetness. The high-resolution structure and the docking model in this study may provide a structural basis to understand the flavour mechanism induced by the sweet protein. •Structures of brazzein variants D29K and R43A were determined by X-ray crystallography to high resolution.•Brazzein variants exhibit significantly altered electrostatic distributions on the surface.•Hypothetical model of each taste receptor T1R2 and T1R3 was utilized from the AlphaFold Protein Structure Database.•Brazzein is bound between the two CRD regions of T1R2 and T1R3 in the docking model.•Altered interaction and energy in the docking model is consistent with the sweetness change.