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Electron microscope autoradiography was used to study the cellular localization of seven glycoproteins rapidly cleared from the circulating plasma of rats and taken up by the liver. I and 15 min after intravenous administration of the 125I-glycoproteins, livers were fixed in situ by perfusion and processed for autoradiography. Autoradiographic grains in the developed sections were found to represent the intact 125 I-ligand. A quantitative analysis of the distribution and concentration (density) of autoradiographic grains over the three major cell types of the liver was then performed. Three molecules, asialo-fetuin, asialo-orosomucoid, and lactosaminated RNase A dimer, the oligosaccharide chains of which terminate in galactose residues, were bound and internalized almost exclusively (>900 /0) by hepatocytes. Conversely, four molecules, the oligosaccharide chains of which terminate in either N-acetyl-glucosamine (agalacto-orosomucoid) or mannose (ahexosamino-orosomucoid, preputial,8-glucuronidase, and mannobiosaminated RNase A dimer), were specifically bound and internalized by cells lining the blood sinusoids-that is, by Kupffer cells and endothelial cells. Endothelial cells were two to six times more active (on a cell volume basis) than were Kupffer cells in the internalization of these four 125 I-ligands. Mannose and N-acetylglucosamine-terminated glycoproteins competed with each other for uptake into either endothelial cells or Kupffer cells, indicating that a single system recognized mannose or Nacetyl-glucosamine residues. Finally, agalacto-orosomucoid and ahexosaminoorosomucoid were also associated with hepatocytes, but competition experiments utilizing excess asialo-orosomucoid demonstrated that residual galactosyl residues were responsible for this association.J. CELL BIOLOGY

The cross-linking reaction between diimido esters and ribonuclease has been studied in terms of the yield of cross-linked dimer with optimum activity toward double-stranded RNA. With dimethyl suberimidate the most satisfactory conditions were condensation for 15 min at pH 7.5-8.0 at 21 degrees C with 1.25 mol equiv of the diimido ester and a protein concentration of 6%. The dimer (yield 20%) had 19 unmodified NH2 groups out of a theoretical 20 for a molecule in which two such groups are involved in the cross-linkage; the activity toward poly(A)-poly(U) in 0.14 M salt solution by spectrophotometric assay was 8.5 times that of the monomeric enzyme toward the same substrate.

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