Purification and characterization of a galactose-specific lectin from corn (Zea mays) coleoptile.
We purified and characterized a lectin from the corn coleoptyle (Zea mays). The lectin (CCL) was purified by affinity chromatography on a Lactosyl-Sepharose 4B column. It is a glycoprotein of 88.7 kDa, composed mainly by glutamic, aspartic, glycine, and Ser residues; in a minor proportion, it contained methionine and cysteine residues. Carbohydrates that constituted 12% of the total weight comprised galactose, mannose, and N-acetyl-D-glucosamine. The lectin contained the blocked amino-terminus. Analysis of the lectin, determined from peptides obtained after trypsin digestion by MALDI-TOF (matrix-assisted laser desorption ionization-time of flight), indicated that CCL has 18% homology with a putative calcium-dependent Ser/Thr protein kinase, from Arabidopsis thaliana, and 39% homology with a NADPH-dependent reductase from Z. mays. The lectin showed hemagglutinating activity toward several erythrocytes, including human A, B, and O. Hapten inhibition assays indicated that the lectin interacts specifically with the OH on C4 from galactose residues. OH- on C1 plays a relevant role in the interaction with CCL, since beta-galactose residues are better recognized than those from the anomeric alpha-galactose. Lack of lectin activity was observed in corn extracts; the highest specific activity was obtained from coleoptyle obtained at the 7th day after seeding.
Martínez-Cruz
M
M
Laboratorio de Bioquímical/Inmunología, Instituto Tecnológico Regional de Oaxaca, Mexico.
Zenteno
E
E
Córdoba
F
F
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D002846:Chromatography, Affinity; D018548:Cotyledon / Q000737:chemistry; D037102:Lectins / Q000737:chemistry / Q000302:isolation & purification*; D008969:Molecular Sequence Data; D010835:Phytohemagglutinins / Q000302:isolation & purification; D037121:Plant Lectins; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013997:Time Factors; D014357:Trypsin; D003313:Zea mays / Q000737:chemistry*
D000596:Amino Acids; D037102:Lectins; D010835:Phytohemagglutinins; D037121:Plant Lectins; D014357:Trypsin