Chemical modification studies on a blood group A-specific lectin, crotalarin (Crotalaria striata) and its effect on hemagglutinating activity.
Crotalarin, the N-acetyl-D-galactosamine-binding blood group A-specific lectin from the seeds of Crotalaria striata was subjected to various chemical modifications in order to ascertain the amino acid residues responsible for its carbohydrate-binding property. Modification of lysine, cysteine and arginine residues did not affect the carbohydrate-binding activity of the lectin. However, modification of tyrosine residue and carboxy group of the acidic amino acids led to a complete loss of its activity, indicating the involvement of tyrosine and aspartic and glutamic acid in the saccharide-binding respectively. The hemagglutinating activity of the lectin was completely/almost completely lost by modification of tryptophan residues. The relative loss in hemagglutinating activity on modification of tryptophan residues indicate that one residue/molecule is required for the carbohydrate-binding activity of the lectin. Modification was not effective in the presence of D-galactose (0.2 M). A marked decrease in the fluorescence emission was found as the tryptophan residues of crotalarin were modified. The c.d. spectra showed the presence of an identical pattern of conformation in the native and modified lectins which confirms that the loss in activity was due to modification only. The effect of periodate oxidation on crotalarin showed loss of activity whereas action of enzymes retained most of the activity.
Sikdar
S
S
Department of Biological Chemistry, Indian Association for the Cultivation of Science, Jadavpur, Calcutta.
Chatterjee
B P
BP
D000017:ABO Blood-Group System; D000107:Acetylation; D000596:Amino Acids / Q000032:analysis; D002236:Carbohydrate Conformation; D002942:Circular Dichroism; D005453:Fluorescence; D006384:Hemagglutination / Q000187:drug effects*; D006386:Hemagglutination Tests; D037102:Lectins / Q000737:chemistry* / Q000494:pharmacology; D010504:Periodic Acid / Q000378:metabolism; D037121:Plant Lectins; D010944:Plants / Q000032:analysis; D013329:Structure-Activity Relationship
D000017:ABO Blood-Group System; D000596:Amino Acids; D037102:Lectins; D037121:Plant Lectins; D010504:Periodic Acid