Carbohydrate specificity of a lectin isolated from the fungus Sclerotium rolfsii.
In order to investigate the functional roles of a phytopathogenic fungal lectin (SRL) isolated from the bodies of Sclerotium rolfsii, the binding properties of SRL were studied by enzyme linked lectinosorbent assay and by inhibition of SRL-glycan interaction. Among glycoproteins (gp) tested for binding, SRL reacted strongly with GalNAc alpha1-->4Ser/Thr (Tn) and/or Gal beta1-->3GalNAc alpha1-->(T(alpha)) containing gps: human T(alpha) and Tn glycophorin, asialo salivary gps, and asialofetuin, but its reactivity toward sialylated glycoproteins was reduced significantly. Of the sugar ligands tested for inhibition of SRL-asialofetuin binding, Thomsen-Friedenreich residue (T(alpha)) was the best, being 22.4 and 2.24 x 10(3) more active than GalNAc and Gal beta1--> residues, respectively. Other ligands tested were inactive. When the glycans used as inhibitors, T(alpha), and/or Tn containing gps, especially asialo PSM, asialo BSM, asialo OSM, active antifreeze gp, asialo glycophorin and Tn-glycophorin were very active, and 1.0 x 10(4) times more potent than GalNAc. From these results, it is clear that the combining site of SRL should be of a cavity type and recognizes only Tn and T(alpha) residues of glycans; it is suggested that T(alpha) and Tn glycotopes, which are present only in abnormal carbohydrate sequences of higher orders of mammal, are the most likely sites for phytopathogenic fungal attachment as an initial step of infection. The affinity of SRL for ligands can be ranked in decreasing order as follows: multivalent T(alpha) and Tn >> monomeric T(alpha) and Tn > GalNAc >>> II (Gal beta1-->4GlcNAc), L (Gal beta1-->4Glc), and Gal.
Wu
A M
AM
Glyco-Immunochemistry Research Laboratory, Institute of Molecular & Cellular Biology, College of Medicine, Chang-Gung University, Kwei-San, Tao-Yuan, Taiwan. amwu@mail.cgu.edu.tw
Wu
J H
JH
Tsai
M S
MS
Hegde
G V
GV
Inamdar
S R
SR
Swamy
B M
BM
Herp
A
A
D000818:Animals; D001203:Ascomycota / Q000378:metabolism*; D050260:Carbohydrate Metabolism; D002241:Carbohydrates / Q000737:chemistry; D006801:Humans; D037102:Lectins / Q000302:isolation & purification / Q000378:metabolism*; D011485:Protein Binding; D013379:Substrate Specificity
D002241:Carbohydrates; D037102:Lectins