Photoaffinity labeling of an acorn barnacle lectin with a photoactivatable fluorescent reagent derivative of D-galactosamine.
A photoactivatable D-galactosamine derivative was prepared by reaction of the amino group of D-galactosamine with 1-azide-5-naphthalene sulfonyl chloride (ANS-Cl). The derivative (GalN-ANS) inhibited the agglutination activity of an acorn barnacle lectin against rabbit erythrocytes to the same extent as D-galactosamine. We used GalN-ANS for photoaffinity labeling of the lectin. The photolabeled lectin was digested with pronase and the digest was separated by reversed-phase high-performance liquid chromatography by monitoring fluorescence and uv absorption to isolate the peptide labeled with GalN-ANS. Amino acid analyses of the labeled peptides revealed that GalN-ANS preferentially covalently labeled two regions in the carbohydrate recognition domain of the lectin. One of them was the highly conserved amino-acid sequence region throughout all calcium-dependent animal lectins.
Muramoto
K
K
School of Fisheries Sciences, Kitasato University, Iwate, Japan.
Kamiya
H
H
D000345:Affinity Labels / Q000138:chemical synthesis / Q000494:pharmacology / Q000528:radiation effects; D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D002846:Chromatography, Affinity; D002851:Chromatography, High Pressure Liquid; D005688:Galactosamine / Q000031:analogs & derivatives / Q000138:chemical synthesis / Q000494:pharmacology / Q000528:radiation effects; D006386:Hemagglutination Tests; D037102:Lectins / Q000032:analysis* / Q000378:metabolism; D008969:Molecular Sequence Data; D009282:Naphthalenesulfonates / Q000138:chemical synthesis / Q000494:pharmacology / Q000528:radiation effects; D010782:Photolysis; D037121:Plant Lectins; D011817:Rabbits; D014466:Ultraviolet Rays
D000345:Affinity Labels; D037102:Lectins; C064593:Megabalanus rosa lectin; D009282:Naphthalenesulfonates; D037121:Plant Lectins; C075247:N-(1-azido-5-naphthalenesulfonyl)galactosamine; D005688:Galactosamine