Isolation and characterisation of a Salvia bogotensis seed lectin specific for the Tn antigen.
A lectin was isolated and characterised from Salvia bogotensis seeds. Removal of the abundant pigments and polysaccharides, which are present in seeds, was an essential step in its purification. Several procedures were assayed and the best suited, including Pectinex treatment, DEAE-cellulose and affinity chromatography, led to a protein being obtained amounting to 18-20mg/100g seeds having high specific agglutination activity (SAA). The lectin specifically agglutinated human Tn erythrocytes and was inhibited by 37mM GalNAc, 0.019mM ovine submaxillary mucin (OSM) or 0.008mM asialo bovine submaxillary mucin (aBSM). Enzyme-linked lectinosorbent assay (ELLSA) revealed strong binding to aOSM and aBSM, corroborating Tn specificity, whereas no binding to fetuin or asialo fetuin was observed. The lectin's monomer MW (38,702Da), amino acid composition, pI, carbohydrate content, deglycosylated form MW, thermal stability and Ca(2+) and Mn(2+) requirements were determined. Evidence of the existence of two glycoforms was obtained. The lectin's specificity and high affinity for the Tn antigen, commonly found in tumour cells, makes this protein a useful tool for immunohistochemical and cellular studies.
Vega
Nohora
N
Biochemistry Laboratory, Chemistry Department, Universidad Nacional de Colombia, Bogotá, Colombia.
Pérez
Gerardo
G
D000116:Acetylgalactosamine / Q000378:metabolism; D000818:Animals; D015295:Antigens, Tumor-Associated, Carbohydrate / Q000378:metabolism*; D001212:Asialoglycoproteins / Q000378:metabolism; D002118:Calcium / Q000494:pharmacology; D002240:Carbohydrate Sequence; D002417:Cattle; D006384:Hemagglutination; D006801:Humans; D006863:Hydrogen-Ion Concentration; D037102:Lectins / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D008345:Manganese / Q000494:pharmacology; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009077:Mucins / Q000378:metabolism; D010936:Plant Extracts / Q000737:chemistry; D027544:Salvia / Q000737:chemistry*; D012639:Seeds / Q000737:chemistry*; D013363:Submandibular Gland / Q000378:metabolism; D013696:Temperature; D014168:Transferrin / Q000031:analogs & derivatives / Q000378:metabolism; D000509:alpha-Fetoproteins / Q000378:metabolism
D015295:Antigens, Tumor-Associated, Carbohydrate; D001212:Asialoglycoproteins; D037102:Lectins; D009077:Mucins; D010936:Plant Extracts; C040286:Tn antigen; D014168:Transferrin; D000509:alpha-Fetoproteins; C017031:asialotransferrins; D008345:Manganese; D000116:Acetylgalactosamine; D002118:Calcium