Comparison of desialylation of rat transferrin by cellular and non-cellular methods.
We have previously shown that the liver endothelium can desialylate the glycoprotein transferrin (Tf). In the present work we provide evidence that asialotransferrin obtained by this means behaves differently on Ricinus communis agglutinin (RCA120) lectin affinity chromatography from asialotransferrin obtained by either neuraminidase treatment or acid hydrolysis. Purified rat transferrin was radiolabelled either with 125I (protein moiety) or with 3H (sialyl residues), and subsequently saturated with iron. It was then passed through an RCA120-agarose column to isolate the fully sialylated component. Sialylated Tf was then desialylated either by incubation with purified rat liver endothelium or, in vitro, by neuraminidase treatment or by acid hydrolysis. The protein was again subjected to RCA120 column chromatography. Although both neuraminidase treatment and acid hydrolysis almost completely desialylated the glycoprotein (as evidenced by near absence of 3H label), the glycoprotein was not retained by the RCA120-agarose column. By contrast, liver endothelium partially desialylated the glycoprotein, but this desialylated fraction was retained by the RCA120-agarose column. These results suggest that desialylation with neuraminidase or acid hydrolysis may be inadequate for functional studies of asialotransferrin.
Irie
S
S
University of Mississippi School of Medicine, Veterans Administration Medical Center, Jackson 39216.
Minguell
J J
JJ
Tavassoli
M
M
D000818:Animals; D001212:Asialoglycoproteins; D002367:Ricinus communis; D002846:Chromatography, Affinity; D002847:Chromatography, Agarose; D004727:Endothelium / Q000378:metabolism; D006868:Hydrolysis; D037102:Lectins; D008099:Liver / Q000378:metabolism; D009439:Neuraminidase; D037121:Plant Lectins; D010947:Plants, Toxic; D051381:Rats; D013464:Sulfuric Acids; D014168:Transferrin / Q000031:analogs & derivatives* / Q000302:isolation & purification
D001212:Asialoglycoproteins; D037102:Lectins; D037121:Plant Lectins; C017707:Ricinus communis agglutinin-1; D013464:Sulfuric Acids; D014168:Transferrin; C017031:asialotransferrins; D009439:Neuraminidase