A comparative study on the purification of the Amaranthus leucocarpus syn. hypocondriacus lectin.
Amaranthus leucocarpus lectin is a homodimeric glycoprotein of 35 kDa per sub-unit, which interacts specifically with N-acetyl-galactosamine. In this work, we compared different glycoproteins that contain Galbeta1-3 GalNAcalpha1-3 Ser/Thr or GalNAcalpha1-3 Ser/Thr in their structure as ligands to purify the A. leucocarpus lectin. From the glycoproteins tested, fetuin was the most potent inhibitor of the hemagglutinating activity and the better ligand for lectin purification; however, the use of desialylated stroma from erythrocytes represented the cheapest method to purify this lectin. O-linked glycans released from the glycoproteins used as affinity matrix and those from different erythrocytes were less inhibitory than parental glycoproteins. The NH2-terminal of the lectin is blocked; moreover, this is the only example of a lectin isolated from this genus to be a glycoprotein. Analysis of the glycoprotein sequences with inhibitory activity for the lectin, showed a different pattern in the O-glycosylation, which confirms that A. leucocarpus lectin recognizes conformation and, probably, distances among O-linked glycans moieties.
Hernández
P
P
Departamento de Bioquímica, Instituto Nacional de Enfermedades Respiratorias, Tlalpan DF, Mexico.
Bacilio
M
M
Porras
F
F
Juarez
S
S
Debray
H
H
Zenteno
E
E
Ortiz
B
B
D000595:Amino Acid Sequence / Q000235:genetics; D000818:Animals; D015295:Antigens, Tumor-Associated, Carbohydrate / Q000737:chemistry* / Q000378:metabolism; D000957:Antigens, Viral, Tumor / Q000737:chemistry* / Q000378:metabolism; D002241:Carbohydrates / Q000494:pharmacology; D002845:Chromatography / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel; D004910:Erythrocyte Membrane / Q000737:chemistry / Q000378:metabolism; D006023:Glycoproteins / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism / Q000494:pharmacology*; D006031:Glycosylation; D006384:Hemagglutination / Q000187:drug effects*; D006801:Humans; D037102:Lectins / Q000302:isolation & purification* / Q000494:pharmacology*; D008969:Molecular Sequence Data; D009077:Mucins / Q000032:analysis; D037121:Plant Lectins; D011134:Polysaccharides / Q000494:pharmacology; D011487:Protein Conformation; D011817:Rabbits; D012639:Seeds / Q000737:chemistry*
C121359:Amaranthus leucocarpus lectin; D015295:Antigens, Tumor-Associated, Carbohydrate; D000957:Antigens, Viral, Tumor; D002241:Carbohydrates; D006023:Glycoproteins; D037102:Lectins; D009077:Mucins; D037121:Plant Lectins; D011134:Polysaccharides; C040286:Tn antigen