Chemical modification studies on a lectin from Saccharomyces cerevisiae (baker's yeast).
The effect of chemical modification on a galactose-specific lectin isolated from a fatty acid auxotroph of Saccharomyces cerevisiae was investigated in order to identify the type of amino acids involved in its agglutinating activity. Modification of 50 free amino groups with succinic anhydride or citraconic anhydride led to an almost complete loss of activity. This could not be protected by the inhibitory sugar methyl alpha-D-galactopyranoside. Treatment with N-bromosuccinimide and N-acetylimidazole, for the modification of tryptophan and tyrosine residues, did not affect lectin activity. Modification of carboxy groups with glycine ethyl ester greatly affected lectin activity, although sugars afford partial protection. Modification of four thiol groups with N-ethylmaleimide was accompanied by a loss of 85% of the agglutinating activity, and two thiol groups were found to be present at the sugar-binding site of the lectin. Modification of 18 arginine residues with cyclohexane-1,2-dione and 26 histidine residues with ethoxyformic anhydride led to a loss of lectin activity. However, in these cases, modification was not protected by the abovementioned inhibitory sugar, suggesting the absence of these groups at the sugar-binding site. In all the cases, immunodiffusion studies with modified lectin showed no gross structural changes which could disrupt antigenic sites of the lectin.
D000596:Amino Acids / Q000032:analysis; D001120:Arginine; D001665:Binding Sites; D002241:Carbohydrates / Q000032:analysis; D055598:Chemical Phenomena; D002621:Chemistry; D004047:Diethyl Pyrocarbonate / Q000494:pharmacology; D006639:Histidine; D005779:Immunodiffusion; D037102:Lectins; D012441:Saccharomyces cerevisiae / Q000032:analysis* / Q000187:drug effects; D013329:Structure-Activity Relationship; D013387:Succinic Anhydrides / Q000494:pharmacology; D013438:Sulfhydryl Compounds
D000596:Amino Acids; D002241:Carbohydrates; D037102:Lectins; D013387:Succinic Anhydrides; D013438:Sulfhydryl Compounds; D006639:Histidine; C031801:succinic anhydride; D001120:Arginine; D004047:Diethyl Pyrocarbonate