Kinetic analyses of the binding of leukemia inhibitory factor to receptor on cells and membranes and in detergent solution.
The equilibrium and kinetic properties of leukemia inhibitory factor (LIF) binding to a range of cell types have been compared. When binding was examined at 4 degrees C, the majority of cells were found to express a single class of high affinity LIF receptor (KD = 20-100 pM; ka = 2-8 x 10(8) min-1 M-1; kd = 0.0004-0.0011 min-1). In contrast, certain activated macrophage populations expressed apparently independent classes of high and low affinity LIF receptor. The low affinity receptors differed from the high affinity receptors in terms of the dissociation rate of the receptor-ligand complex (KD = 1-2 nM; ka = 3-7 x 10(8) min-1 M-1; kd = 0.30-0.67 min-1). At 37 degrees C, the interaction of LIF with its high affinity receptor was more complicated, since occupied LIF receptors were internalized more rapidly than unoccupied receptors, internalized LIF was hydrolyzed and released from the cell, and new receptors were synthesized and expressed on the cell surface. Interestingly, when membranes were prepared from cells that expressed only high affinity receptors, both high and low affinity receptors were detected, while after detergent solubilization of membranes only low affinity receptors were apparent. These results are discussed in terms of a structural model for the LIF receptor in which interaction of a low affinity binding subunit and a second nonbinding subunit is required for the generation of the high affinity receptor.
Hilton
D J
DJ
Walter and Eliza Hall Institute for Medical Research, Parkville, Victoria, Australia.
Nicola
N A
NA
D016475:3T3 Cells; D000818:Animals; D003001:Cloning, Molecular; D003902:Detergents; D006131:Growth Inhibitors / Q000378:metabolism*; D015850:Interleukin-6; D007457:Iodine Radioisotopes; D007700:Kinetics; D053605:Leukemia Inhibitory Factor; D053680:Leukemia Inhibitory Factor Receptor alpha Subunit; D008222:Lymphokines / Q000235:genetics / Q000378:metabolism*; D008264:Macrophages / Q000378:metabolism; D051379:Mice; D018121:Receptors, Cytokine; D011971:Receptors, Immunologic / Q000378:metabolism*; D053606:Receptors, OSM-LIF; D012996:Solutions; D013379:Substrate Specificity; D014407:Tumor Cells, Cultured
D003902:Detergents; D006131:Growth Inhibitors; D015850:Interleukin-6; D007457:Iodine Radioisotopes; D053605:Leukemia Inhibitory Factor; D053680:Leukemia Inhibitory Factor Receptor alpha Subunit; C507838:Lif protein, mouse; C507841:Lifr protein, mouse; D008222:Lymphokines; D018121:Receptors, Cytokine; D011971:Receptors, Immunologic; D053606:Receptors, OSM-LIF; D012996:Solutions