The isolation of a rat alveolar macrophage lectin.
A lectin in rat alveolar macrophage membranes with a high affinity for binding ligands containing L-fucose and N-acetyl-D-glucosamine has been isolated by affinity chromatography on Fuc-BSA-Sepharose (where Fuc is fucosyl and BSA is bovine serum albumin). The lectin was extracted from rat lung homogenates with Triton X-100, absorbed from the extract onto Fuc-BSA-Sepharose in the presence of Ca2+ and eluted by removal of Ca2+. After a second adsorption to and elution from Fuc-BSA-Sepharose, three protein species were detected electrophoretically in fractions that bind Fuc-BSA. One, which was the mannose/N-acetylglucosamine lectin (Mr = 32,000) found earlier in hepatocytes, was removed by adsorption on anti-lectin IgG-Sepharose. Another (Mr = 46,000) was removed by adsorption to Fuc-BSA-Sepharose and elution with galactose. The remaining lectin (Mr = 180,000) bound fucose and N-acetylglucosamine but not galactose. Binding was maximal between pH 6.5 and 9.0 and dependent on Ca2+. Immunocytological analysis with rabbit anti-lectin IgG and fluorescein-labeled goat anti-rabbit IgG revealed the lectin to be in rat alveolar macrophages and nonparenchymal cells of liver. Thus, the lectin appears to be present in macrophages and is likely involved in receptor-mediated endocytosis. It is distinctly different structurally from the hepatocyte lectin with a similar ligand-binding specificity.
R S
RS
Haltiwanger
R L
RL
Hill
D000818:Animals; D000906:Antibodies / Q000276:immunology; D002118:Calcium / Q000494:pharmacology; D002846:Chromatography, Affinity; D005643:Fucose / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D066298:In Vitro Techniques; D037102:Lectins / Q000276:immunology / Q000302:isolation & purification*; D008099:Liver / Q000032:analysis; D008264:Macrophages / Q000032:analysis*; D008297:Male; D008970:Molecular Weight; D011650:Pulmonary Alveoli / Q000032:analysis*; D011817:Rabbits; D051381:Rats; D011919:Rats, Inbred Strains; D012710:Serum Albumin, Bovine / Q000378:metabolism
D000906:Antibodies; D037102:Lectins; D012710:Serum Albumin, Bovine; D005643:Fucose; D002118:Calcium