Highly probable active site of the sweet protein monellin.
The sweet protein monellin consists of two noncovalently associated polypeptide chains, the A chain of 44 amino acid residues and the B chain of 50 residues. Synthetic monellin is 4000 times as sweet as sucrose on a weight basis, and the native conformation is essential for the sweet taste. Knowledge of the active site of monellin will provide important information on the mode of interaction between sweeteners and their receptors. If the replacement of a certain amino acid residue in monellin removes the sweet taste, while the native conformation is retained, it may be concluded that the position replaced is the active site. Our previous replacement studies on Asp residues in the A chain did not remove the sweet taste. The B chain contains two Asp residues at positions 7 and 21, which were replaced by Asn. [AsnB21]Monellin and [AsnB7]monellin were 7000 and 20 times sweeter than sucrose, respectively. The low potency of the [AsnB7]monellin indicates that AspB7 participates in binding with the receptor. AspB7 was then replaced by Abu. [AbuB7]Monellin was devoid of sweetness, and retained the native conformation. AspB7 is located at the surface of the molecule (Ogata et al.). These results suggest that Asp7 in the B chain is the highly probable active site of monellin.
Central Research Laboratories, Ajinomoto Co., Inc., Kawasaki, Japan.
M
M
Kohmura
N
N
Nio
Y
Y
Ariyoshi
D000595:Amino Acid Sequence; D001665:Binding Sites; D002851:Chromatography, High Pressure Liquid; D002942:Circular Dichroism; D007202:Indicators and Reagents; D046911:Macromolecular Substances; D008969:Molecular Sequence Data; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry* / Q000302:isolation & purification; D010940:Plant Proteins / Q000138:chemical synthesis / Q000737:chemistry*; D011487:Protein Conformation; D013050:Spectrometry, Fluorescence; D013329:Structure-Activity Relationship; D014675:Vegetables / Q000737:chemistry*
D007202:Indicators and Reagents; D046911:Macromolecular Substances; D010455:Peptides; D010940:Plant Proteins; C002541:monellin protein, Dioscoreophyllum cumminsii