Binding interactions of leukemia inhibitory factor and ciliary neurotrophic factor with the different subunits of their high affinity receptors.
Leukemia inhibitory factor (LIF) and ciliary neurotrophic factor (CNTF) share common components in their multimeric receptors. Both cytokine receptors contain gp130/interleukin-6-receptor transducer as well as gp190/low affinity LIF receptor. For CNTF, addition of a third subunit, or alpha subunit, defines the high-affinity CNTF receptor. In the present study, we analyzed the binding interactions of LIF and CNTF in human cell lines and showed a mutual displacement for LIF and CNTF toward the trimeric high-affinity CNTF receptor. Similar results were obtained in the JEG cell line, which only expressed the gp130/gp190 high-affinity LIF receptor, by adding a soluble form of the alpha CNTF receptor to the system to reconstitute the high-affinity-type CNTF receptor. The different receptor subunits were then expressed separately in transfected cells and their binding capacities analyzed. The results showed that the heterocomplex CNTF/alpha CNTF receptor bound to gp130 with an affinity of 3-5 x 10(-10)M, whereas LIF interacted mainly with gp190. In summary, the observed competition between LIF and CNTF does not result from the binding to a common site or receptor subunit, but rather to the interaction of the three receptor components to create a conformational site common to both LIF and CNTF.
Robledo
O
O
Laboratoire de Biologie Cellulaire, CHU Angers, France.
Auguste
P
P
Coupey
L
L
Praloran
V
V
Chevalier
S
S
Pouplard
A
A
Gascan
H
H
D000818:Animals; D001667:Binding, Competitive / Q000502:physiology; D002522:Chlorocebus aethiops; D002822:Choriocarcinoma; D020934:Ciliary Neurotrophic Factor; D006131:Growth Inhibitors / Q000378:metabolism*; D006801:Humans; D015850:Interleukin-6; D053605:Leukemia Inhibitory Factor; D053680:Leukemia Inhibitory Factor Receptor alpha Subunit; D008222:Lymphokines / Q000378:metabolism* / Q000648:ultrastructure; D009414:Nerve Growth Factors / Q000378:metabolism*; D009419:Nerve Tissue Proteins / Q000378:metabolism*; D009447:Neuroblastoma; D011485:Protein Binding / Q000502:physiology; D011869:Radioligand Assay; D020801:Receptor, Ciliary Neurotrophic Factor; D018121:Receptors, Cytokine / Q000378:metabolism* / Q000648:ultrastructure; D017475:Receptors, Nerve Growth Factor / Q000378:metabolism* / Q000648:ultrastructure; D053606:Receptors, OSM-LIF; D013997:Time Factors; D014407:Tumor Cells, Cultured / Q000378:metabolism / Q000648:ultrastructure
D020934:Ciliary Neurotrophic Factor; D006131:Growth Inhibitors; D015850:Interleukin-6; C507837:LIF protein, human; C507840:LIFR protein, human; D053605:Leukemia Inhibitory Factor; D053680:Leukemia Inhibitory Factor Receptor alpha Subunit; D008222:Lymphokines; D009414:Nerve Growth Factors; D009419:Nerve Tissue Proteins; D020801:Receptor, Ciliary Neurotrophic Factor; D018121:Receptors, Cytokine; D017475:Receptors, Nerve Growth Factor; D053606:Receptors, OSM-LIF