The chromatographic heterogeneity of rat transferrin on immobilized concanavalin A and lentil lectin.
A procedure was developed for the isolation of the microheterogeneous forms of rat transferrin consisting of anion-exchange and serial lectin affinity chromatographies. By deploying this technique, four to five different anionic species of the protein were detected in plasma. The two major components obtained, which encompassed 92-94% of the plasma transferrin, were further studied by sequential lectin chromatography. The larger of the two, representing 60-63% of plasma transferrin, was bound by concanavalin A - Sepharose, while the smaller one (30-32% of plasma transferrin) resolved into an unbound (25-27% of plasma transferrin) and a retarded (4-5% of plasma transferrin) fraction. The latter eluted from the column in a volume which was 1.9 times larger than that required for the passage of nonretarded transferrin. In accordance with their fucose contents, each of these three concanavalin A fractions resolved into a bound (20-29%) and an unbound (71-80%) subfraction by chromatography on lentil-Sepharose. It is concluded that there exist two kinds of glycan microheterogeneity in rat transferrin and that they are unrelated to each other. Consequently, at least six different forms of rat transferrin are available with respect to glycosylation. Epididymal fucosidase cleaved fucose from apotransferrin slowly and from the tryptic glycopeptide rapidly. Exploratory studies performed in vivo failed thus far to identify the significance of fucose in rat transferrin.
Department of Pathology, McMaster University Health Sciences Center, Hamilton, Ont., Canada.
E
E
Regoeczi
P A
PA
Chindemi
J R
JR
Rudolph
G
G
Spik
J
J
Montreuil
D000818:Animals; D002846:Chromatography, Affinity; D002852:Chromatography, Ion Exchange; D003208:Concanavalin A / Q000378:metabolism; D005643:Fucose / Q000032:analysis; D037102:Lectins / Q000378:metabolism; D037121:Plant Lectins; D051381:Rats; D013329:Structure-Activity Relationship; D014168:Transferrin / Q000032:analysis* / Q000378:metabolism
D037102:Lectins; D037121:Plant Lectins; D014168:Transferrin; C033619:lentil lectin; D003208:Concanavalin A; D005643:Fucose