Purification and characterization of the D-mannose receptor from J774 mouse macrophage cells.
Macrophages display on their cell surface a D-mannose-specific receptor which facilitates the scavenging of certain pathogens and deleterious macromolecules from the extracellular fluid as part of the host defense mechanism. The mouse D-mannose receptor was purified from J774 E macrophages and an antiserum was generated against the receptor protein. In mouse macrophages, the newly synthesized receptor has an Mr of 157,000 Da and rapidly matures to a protein with an Mr of 172,000 Da. Both forms of the receptor protein are tightly associated with cell membranes. The receptor is found in a number of mouse macrophage cell types but is not present in mouse fibroblasts. An assay was developed to characterize D-mannose receptor-ligand binding based on immunoprecipitation of the detergent-solubilized receptor protein. The dissociation constant, determined for receptor and the neoglycoprotein D-mannose-BSA, was 1.67nM. Receptor-ligand binding was calcium and pH dependent. Monosaccharides, such as D-mannose and L-fucose, partially inhibited receptor binding to the ligand D-mannose-BSA.
Department of Cell Biology and Physiology, Washington University School of Medicine, St. Louis, Missouri 63110.
J S
JS
Blum
P D
PD
Stahl
R
R
Diaz
M L
ML
Fiani
D000818:Animals; D002460:Cell Line; D006863:Hydrogen-Ion Concentration; D037181:Lectins, C-Type; D008264:Macrophages / Q000378:metabolism*; D008358:Mannose / Q000378:metabolism*; D000090323:Mannose Receptor; D037241:Mannose-Binding Lectins; D051379:Mice; D008970:Molecular Weight; D011956:Receptors, Cell Surface; D011971:Receptors, Immunologic / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism
D037181:Lectins, C-Type; D000090323:Mannose Receptor; D037241:Mannose-Binding Lectins; D011956:Receptors, Cell Surface; D011971:Receptors, Immunologic; D008358:Mannose