Isolation and characterization of a soluble lactose-binding lectin from postnatal chicken retina.
We investigated the presence of endogenous lectins in postnatal chicken retinal tissue assaying the hemagglutinating activity of crude soluble extracts of the tissue that was homogenized in a buffer supplemented with different sugars. Lactose was the most effective sugar to extract an hemagglutinating activity. Using similar extraction conditions, other sugars, such as glucose, N-acetylglucosamine, mannose, fucose, glucuronic and sialic acid, were ineffective to extract any significant hemagglutinating activity. The lectin was purified by affinity chromatography on lactosyl-Sepharose. SDS-PAGE and isoelectric focusing analyses showed that it has a subunit molecular weight of 16 kDa and a pI about 4.5. The retinal lectin cross-reacted immunologically with a rabbit antiserum raised against a lectin purified from adult chicken liver, which is a CLL-I (Beyer et al.: J Biol Chem 255:4236-4239, 1980) or C-16 (Sakakura et al.: J Biol Chem 265:21573-21579, 1990) form of chicken endogenous soluble lactose-binding lectins. Gel filtration studies showed that the oligomeric structure of the retinal lectin is dependent on the ionic strength of the elution buffer. The lectin hemagglutinating activity and the amount of lectin protein reached their highest levels at late developmental stages of the retinal tissue, suggesting that retinal lectin might have a functional role during terminal differentiation of retinal cells.
Castagna
L F
LF
Centro de Investigaciones en Química Biológica de Córdoba, Facultad de Ciencias Químicas, Universidad Nacional de Córdoba, Argentina.
Landa
C A
CA
D000818:Animals; D002645:Chickens; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D003429:Cross Reactions; D004591:Electrophoresis, Polyacrylamide Gel; D037541:Galectin 4; D006386:Hemagglutination Tests; D006388:Hemagglutinins / Q000302:isolation & purification*; D015151:Immunoblotting; D007422:Intestines / Q000737:chemistry; D007525:Isoelectric Focusing; D037102:Lectins / Q000302:isolation & purification*; D008024:Ligands; D008099:Liver / Q000737:chemistry; D008970:Molecular Weight; D012160:Retina / Q000737:chemistry*
D037541:Galectin 4; D006388:Hemagglutinins; D037102:Lectins; D008024:Ligands