Purification and some characteristics of a beta-galactoside binding soluble lectin from amphibian ovary.
Soluble extracts of Bufo ovaries agglutinate sialidase-treated rabbit erythrocytes. Unlike other amphibian lectins this agglutination activity does not require the presence of calcium ions. It is specifically inhibited by D-galactose and its derivatives. Thiodi-D-galactoside is the most potent saccharide inhibitor followed by lactose and methyl-beta-D-galactoside, respectively. D-Fucose, D-glucose and D-mannose do not inhibit the activity at concentrations at or above 100 mM. The lectin has been purified 500-fold to apparent homogeneity from the ovaries by salt extraction and affinity chromatography on lactose-aminophenyl-agarose, with a yield of about 0.2%. The molecular mass determined by gel filtration under native conditions was 30 kDa; polyacrylamide gel electrophoresis in SDS gave a molecular mass of 15 kDa, suggesting that the lectin is a dimer. The lectin has an isoelectric point of 40 and contains a high proportion of acidic amino acids.
Fink de Cabutti
N E
NE
Instituto de Biologia de la Reproducion y Desarrollo Embrionario, Universidad Nacional de Lomas de Zamora, Argentina.
Caron
M
M
Joubert
R
R
Elola
M T
MT
Bladier
D
D
Herkovitz
J
J
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002025:Bufonidae; D005260:Female; D005697:Galactosides / Q000378:metabolism*; D006027:Glycosides / Q000378:metabolism*; D006388:Hemagglutinins / Q000302:isolation & purification; D007526:Isoelectric Point; D037102:Lectins / Q000302:isolation & purification*; D008970:Molecular Weight; D010053:Ovary / Q000032:analysis*
D000596:Amino Acids; D005697:Galactosides; D006027:Glycosides; D006388:Hemagglutinins; D037102:Lectins