The interaction of Abrus precatorius agglutinin with saccharides as analyzed by fluorescence spectroscopy.
The binding of saccharides to Abrus precatorius agglutinin (APA) was analyzed by fluorescence spectroscopy. Upon binding of specific saccharides, the fluorescence emission maximum of APA (338 nm) shifted to shorter wavelength by 5 nm, owing to the change in the environment of tryptophan. By analyzing the change in the fluorescence intensity at 338 nm as a function of concentration of saccharides, the association constants for binding of saccharides to APA were determined. The results suggest that in the saccharide binding site on each B-chain of APA, there may be a site which interacts with the saccharide residue linked to galactopyranoside at the non-reducing end, in addition to the site which recognizes the galactopyranosyl residue. Fluorescence quenching data indicate that 8 out of 24 tryptophans in APA are located at or near the surface of the protein molecule and are available for quenching with both KI and acrylamide, and 10 tryptophans are involved in the environment to which acrylamide has access but KI does not. Binding of lactose to APA reduced by 4 the number of tryptophan residues accessible to quenchers. Based on the results, it is suggested that the tryptophan residues at the saccharide binding site on each B-chain of APA are present on the surface of the APA molecule, and they are shielded from quenching by KI and acrylamide upon binding with specific saccharides.
Yamasaki
N
N
Laboratory of Biochemistry, Faculty of Agriculture, Kyushu University, Fukuoka.
Absar
N
N
Funatsu
G
G
D000178:Acrylamides / Q000032:analysis; D002241:Carbohydrates / Q000032:analysis*; D055598:Chemical Phenomena; D002621:Chemistry; D007785:Lactose / Q000032:analysis; D037102:Lectins / Q000032:analysis*; D011193:Potassium Iodide / Q000032:analysis; D013050:Spectrometry, Fluorescence; D014364:Tryptophan / Q000032:analysis
D000178:Acrylamides; D002241:Carbohydrates; D037102:Lectins; D011193:Potassium Iodide; D014364:Tryptophan; D007785:Lactose