Isolation and characterization of soluble β-galactoside-binding lectins from mammalian liver

• Published in: Biochimica et biophysica acta Vol. 992; no. 1; pp. 30 - 34
• Main Authors: Ali, Najma, Salahuddin, A.
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 21.07.1989; Elsevier; North-Holland
• Subjects: Analytical, structural and metabolic biochemistry; Animals; Binding and carrier proteins; Biological and medical sciences; Buffaloes; Characterization; Electrophoresis, Polyacrylamide Gel; Fundamental and applied biological sciences. Psychology; Galactosides - isolation & purification; Galectins; Glycosides - isolation & purification; Goats; Hemagglutination; Hemagglutinins - isolation & purification; Hepatic lectin; Lectin; Liver - metabolism; Proteins; Rabbits; Sheep; Species Specificity; Spectrometry, Fluorescence; Spectrophotometry, Ultraviolet; Sulfhydryl Compounds - antagonists & inhibitors; β-Galactoside binding lectin; PBS; Lectin; β-Galactoside binding lectin; HL; Characterization; SDS-PAGE; ASF; PMSF; NEM; pHMB; RL; R m; Hepatic lectin; Binding protein; Vertebrata; Molecular form; Hemagglutination; Mammalia; Purification; Affinity chromatography; Liver; Sheep; Artiodactyla; Ungulata
• ISSN: 0304-4165; 0006-3002; 1872-8006
• DOI: 10.1016/0304-4165(89)90046-9

Soluble β-galactoside-binding lectins were isolated by chromatography on asialofetuin-Sepharose-4B column in 10 mM Tris-HCl buffer (pH 7.5) containing 150 mM NaCl, 5 mM CaCl 2 and 1 mM 2-mercaptoethanol. The three lectins moved essentially as single polypeptide bands, of 18, 22 and 24 kDa, respectively, for sheep, goat and buffalo hepatic lectins. Sheep and goat lectins each contained 4 mol of hexose, whereas the hexose content of the buffalo lectin was 7 mol. The number of sulfhydryl groups in sheep, goat and buffalo lectins were determined to be 3.2, 4.3 and 4.8, respectively. The optical properties of the three lectins were similar to those of tryptophan-containing proteins. Lectin mediated hemagglutination of trypsinized rabbit erythrocytes was most effectively inhibited by lactose, followed by o-nitrophenyl β-galactopyranoside and galactose, but remained unaffected by glucose, mannose, fucose and fructose. Calcium ions substantially enhanced their hemagglutinating activity. Goat and buffalo lectins, but not sheep lectin, were also stimulated by Mg 2+, Mn 2+, Sr 2+ and Ni 2+ ions. The lectins lost activity after treatment with para-hydroxy-mercuribenzoate and N-ethylmaleimide. However, iodoacetamide treatment had no effect on the activity. The results show that the three lectins are different from the soluble β-galactoside-binding lectins studied thus far.