Abstract
A lectin with a high affinity for binding ligands through fucose residues has been purified to homogeneity from rat liver. Affinity chromatography of the lectin on fucosyl-bovine serum albumin-agarose is the key step in the purification. Contaminating amounts of a previously described lectin that binds mannose and N-acetylglucosamine are removed from the fucose-binding lectin by either immunoadsorption on anti-mannose/N-acetylglucosamine lectin IgG-agarose or by specific elution of the fucose-binding lectin from fucosyl-bovine serum albumin-agarose. The pure fucose-binding lectin contains two polypeptide subunits with molecular weights of 88,000 and 77,000, respectively, as judged by gel electrophoresis. Peptide maps of the subunits, however, show that they are very similar structurally. In addition, peptide maps show that the fucose lectin is structurally distinct from other rat hepatic lectins. This is supported by the lack of cross-reaction among the different rat liver lectins and their specific antibodies and the inability of specific antibodies to the mannose/N-acetylglucosamine lectin to inhibit the binding of fucosyl-bovine serum albumin by the fucose lectin.
Highlights
A lectin with ahigh affinity forbindingligands affinity absorbent, faucose-bindinglectin was separated from through fucose residues has been purified to homoge- the other lectins and shown to be structurally unrelated to neity from rat liver
In order to establish whethera fucose-binding lectinispresent in rat liver that differs from the known hepatic lectins, attempts were made to purify it from liver extracts by affinity chromatography on Fuc-bovine serum albumin (BSA)-agarose.’
Since the original extract of the liver homogenate contained the hepatocyte galactose [21] and the mannose/N-acetylglucosamine-bindinglectins [22] and the latter bound Fuc-BSA [4],it was impossible to estimate the specific activity of the fucose lectin and its yield throughout the purification
Summary
A lectin with ahigh affinity forbindingligands affinity absorbent, faucose-bindinglectin was separated from through fucose residues has been purified to homoge- the other lectins and shown to be structurally unrelated to neity from rat liver. The binding specificity of the lectin is distinctly lectin on fucosyl-bovine serum albumin-agaroissethe different from those of the other known hepatic lectins as key step in the purification. In order to establish whethera fucose-binding lectinispresent in rat liver that differs from the known hepatic lectins, attempts were made to purify it from liver extracts by affinity chromatography on Fuc-BSA-agarose.’. We wish to report here procedures for the purification of a rat liver lectin which has a high binding affinity for certain fucose-containing glycoproteins. The purification of this lectin was complicated by the fact that the mannose/N-acetylglucosamine [2] and the galactose [3] lectins of hepatocytes were present in liver extracts. Present address: Dept. of Molecular Genetics, University of Texas Health Sciences Center at Dallas, 5323 Harry Hines Blvd., Dallas, TX 75235
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