Isolation of a beta-galactoside-binding lectin from cat liver.
A lectin from cat liver has been identified and purified by affinity chromatography on asialofetuin-Sepharose. One hundred micrograms of lectin was obtained from one cat liver with a purification factor of 1561. The lectin agglutinates trypsin-treated rabbit and cow erythrocytes. Hemagglutination was inhibited only by saccharides containing  -galactosyl residues, of which the 1-amine-1-deoxy- -D-galactose was the most potent one by inhibiting hemagglutination at a concentration of 12.5 mM, followed by melibiose, trehalose and galactose. The lectin has a subunit molecular mass of 14.4 kDa determined by SDS-PAGE under reducing conditions and a pI of 4.85. Compared with the composition of lectins from calf heart and porcine heart, cat liver lectin contains approximately the same amount of cysteine, half the amount of glycine, twice as much arginine and threonine, and three times the amounts of tyrosine and methionine. Cat liver lectin contains four cysteine residues per subunit, all of them in the reduced form. Their lack of reactivity towards thiol-reactive supports suggests they are not exposed on the lectin surface. The protein apparently has a blocked N-terminus. The purified lectin was stable for up to 20 months stored at +4 C in buffer supplemented with 4 mM  -mercaptoethanol. Results indicated that this lectin belongs to the family of soluble  -galactoside-binding lectins, also known as galectins, which are expressed in a wide range of vertebrate tissues.
Franco-Fraguas
L
L
Cátedra de Bioquímica, Facultad de Química, Montevideo, Uruguay. lfranco@fq.edu.uy
Batista-Viera
F
F
Carlsson
J
J
D000818:Animals; D002415:Cats; D002417:Cattle; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D004912:Erythrocytes / Q000378:metabolism; D037161:Galectins / Q000737:chemistry / Q000187:drug effects / Q000302:isolation & purification*; D006385:Hemagglutination Inhibition Tests; D008099:Liver / Q000737:chemistry*; D008970:Molecular Weight; D011817:Rabbits; D013439:Sulfhydryl Reagents / Q000494:pharmacology
D037161:Galectins; D013439:Sulfhydryl Reagents