Defining the carbohydrate specificities of aplysia gonad lectin exhibiting a peculiar D-galacturonic acid affinity.
Aplysia gonad lectin (AGL), which has been shown to stimulate mitogenesis in human peripheral lymphocytes, to suppress tumor cells, and to induce neurite outgrowth and improve cell viability in cultured Aplysia neurons, exhibits a peculiar galacturonic acid/galactose specificity. The carbohydrate binding site of this lectin was characterized by enzyme-linked lectino-sorbent assay and by inhibition of AGL-glycan interactions. Examination of the lectin binding with 34 glycans revealed that it reacted strongly with the following glycoforms: most human blood group precursor (equivalent) glycoproteins (gps), two Galalpha1-->4Gal-containing gps, and two d-galacturonic acid (GalUA)-containing polysaccharides (pectins from apple and citrus fruits), but poorly with most human blood group A and H active and sialylated gps. Among the GalUA and mammalian saccharides tested for inhibition of AGL-glycan binding, GalUA mono- to trisaccharides were the most potent ones. They were 8.5 x 10(4) times more active than Gal and about 1.5 x 10(3) more active than the human blood group P(k) active disaccharide (E, Galalpha1-->4Gal). This disaccharide was 6, 28, and 120 times more efficient than Galbeta1-->3GlcNAc(I), Galbeta1-->3GalNAc(T), and Galbeta1--> 4GlcNAc (II), respectively, and 35 and 80 times more active than melibiose (Galalpha1-->6Glc) and human blood group B active disaccharide (Galalpha1-->3Gal), respectively, showing that the decreasing order of the lectin affinity toward alpha-anomers of Gal is alpha1-->4 > alpha1-->6 > alpha1-->3. From the data provided, the carbohydrate specificity of AGL can be defined as GalUAalpha1-->4 trisaccharides to mono GalUA > branched or cluster forms of E, I, and II monomeric E, I, and II, whereas GalNAc is inactive.
Wu
A M
AM
Glyco-Immunochemistry Research Laboratory, Institute of Molecular and Cellular Biology, School of Medicine, Chang-Gung University, Kwei-san 33332, Taiwan. amwu@mail.cgu.edu.tw
Song
S C
SC
Chen
Y Y
YY
Gilboa-Garber
N
N
D000818:Animals; D000941:Antigens / Q000276:immunology / Q000378:metabolism; D001048:Aplysia; D001665:Binding Sites; D001789:Blood Group Antigens / Q000276:immunology; D050260:Carbohydrate Metabolism; D002240:Carbohydrate Sequence; D037161:Galectins; D006388:Hemagglutinins / Q000737:chemistry / Q000378:metabolism*; D006603:Hexuronic Acids / Q000378:metabolism*; D006801:Humans; D008969:Molecular Sequence Data; D011134:Polysaccharides / Q000378:metabolism
D000941:Antigens; D001789:Blood Group Antigens; D037161:Galectins; D006388:Hemagglutinins; D006603:Hexuronic Acids; D011134:Polysaccharides; C007819:galacturonic acid