Possible active site of the sweet-tasting protein thaumatin.
Epitopes on thaumatin and monellin were studied using the PEPSCAN-technology. The antibodies used were raised against thaumatin. Only antibodies that, in an ELISA, both recognized thaumatin and monellin were used in the PEPSCAN-analyses. On thaumatin two major overlapping epitopes were identified. On monellin no epitopes could be identified. The identified epitope region on thaumatin shares structural features with various peptide and protein sweeteners. It contains an aspartame-like site which is formed by Asp21 and Phe80, tips of the two extruding loops KGDAALDAGGR19-29 and CKRFGRPP77-84, which are spatially positioned next to each other. Furthermore, sub-sequences of the KGDAALDAGGR19-29 loop are similar to peptide-sweeteners such as L-Asp-D-Ala-L-Ala-methyl ester and L-Asp-D-Ala-Gly-methyl ester. Since the aspartame-like Asp21-Phe80 site and the peptide-sweetener-like sequences are also not present in non-sweet thaumatin-like proteins it is postulated that the KGDAALDAGGR19-29- and CKRFGRPP77-84 loop contain important sweet-taste determinants. This region has previously not been implicated as a sweet-taste determinant of thaumatin.
Slootstra
J W
JW
Department of Biomolecular Recognition, Institute for Animal Science and Health, Lelystad, The Netherlands.
De Geus
P
P
Haas
H
H
Verrips
C T
CT
Meloen
R H
RH
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D001218:Aspartame / Q000737:chemistry / Q000494:pharmacology; D001665:Binding Sites; D002628:Chemoreceptor Cells / Q000737:chemistry / Q000276:immunology / Q000378:metabolism; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes; D005260:Female; D006825:Hybridomas / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D010940:Plant Proteins / Q000737:chemistry / Q000276:immunology / Q000494:pharmacology*; D011487:Protein Conformation; D013549:Sweetening Agents / Q000737:chemistry / Q000494:pharmacology*; D013649:Taste / Q000187:drug effects*
D000911:Antibodies, Monoclonal; D000939:Epitopes; D010940:Plant Proteins; D013549:Sweetening Agents; C002541:monellin protein, Dioscoreophyllum cumminsii; C003427:thaumatin protein, plant; D001218:Aspartame