Purification of acetyllactosamine-specific tomato lectin by erythroglycan-sepharose affinity chromatography.
Tomato lectin is specific for oligomers of poly-N-acetyllactosamine containing 3 repeating Gal(beta 1-4)GlcNAc (beta 1-3)-disaccharides. As such it is highly useful for purifying oligosaccharides or glycopeptides with poly-N-acetyllactosamine character. We have found the lectin very useful as an affinity reagent for isolating glycoproteins or glycoprotein domains having poly-N-acetyllactosamine glycosylation. Conventional preparation of tomato lectin by ovomucoid-Sepharose affinity chromatography was found to be unsatisfactory due to instability of column and bleeding of ovomucoid into eluents requiring the necessity for additional purification steps following affinity chromatography. We prepared a column of human erythrocyte band 3 carbohydrate glycopeptide (erythroglycan) attached to Sepharose as an affinity matrix. The purification of tomato lectin to homogeneity in one step on this column matrix is described in this report.
Zhu
B C
BC
Department of Biochemistry, Louisiana State University, Baton Rouge.
Laine
R A
RA
D000606:Amino Sugars; D001457:Anion Exchange Protein 1, Erythrocyte; D002846:Chromatography, Affinity; D037102:Lectins / Q000032:analysis / Q000302:isolation & purification*; D008565:Membrane Proteins; D008970:Molecular Weight; D037121:Plant Lectins
D000606:Amino Sugars; D001457:Anion Exchange Protein 1, Erythrocyte; D037102:Lectins; D008565:Membrane Proteins; D037121:Plant Lectins; C018059:erythroglycan; C034981:tomato lectin; C000458:N-acetyllactosamine