Simultaneous localization of an hepatic binding protein specific for galactose and of galactose-containing receptors on rat hepatocytes.
The hepatic binding protein, specific for galactose-terminated glycoproteins (asialoglycoproteins) and the receptors for the Ricinus communis lectin, specific for galactose residues (RCA1), were simultaneously localized on isolated rat hepatocytes by the gold method. The marker for the binding protein was prepared from gold granules (5 nm in diam.) labeled with ceruloplasmin and desialylated. The marker specific for galactose-containing receptors consisted of granules (17 nm in diameter) labeled with RCA1. It was established that both markers did not interact. Hepatocytes (fresh or briefly fixed with glutaraldehyde) were successively incubated with the asialoceruloplasmin and the RCA1 marker. Examination of thin sections by electron microscopy indicated that the binding protein and the RCA1 receptors were often in the proximity of each other on the plasmamembrane. Using the same technique, wheat germ agglutinin (WGA) receptors were generally found on area of the plasmamembrane poorly marked by the RCA1 gold marker. The binding of asialoceruloplasmin gold markers was studied as a function of the size of the granules. It became insignificant when the size was above 17 nm. Previous results have shown that the binding of RCA1 is low when the marker reaches 50 nm in size while WGA markers up to 75 nm are well bound by hepatocytes. It is therefore hypothesized that the binding protein and RCA1 receptors are located between glycoprotein brushes of increasing spacing while part or all of the WGA receptors are located at the periphery of the brushes.
D000818:Animals; D002570:Ceruloplasmin / Q000378:metabolism; D005690:Galactose; D006023:Glycoproteins / Q000378:metabolism; D006046:Gold / Q000378:metabolism; D006801:Humans; D037102:Lectins; D008099:Liver / Q000166:cytology*; D037121:Plant Lectins; D010947:Plants, Toxic; D011485:Protein Binding; D051381:Rats; D012278:Ricinus / Q000378:metabolism; D012568:Schizosaccharomyces / Q000648:ultrastructure; D013997:Time Factors; D014908:Triticum
D006023:Glycoproteins; D037102:Lectins; D037121:Plant Lectins; D006046:Gold; D002570:Ceruloplasmin; D005690:Galactose