Localization of functional receptor epitopes on the structure of ciliary neurotrophic factor indicates a conserved, function-related epitope topography among helical cytokines.
By rational mutagenesis, receptor-specific functional analysis, and visualization of complex formation in solution, we identified individual amino acid side chains involved specifically in the interaction of ciliary neurotrophic factor (CNTF) with CNTFR alpha and not with the beta-components, gp130 and LIFR. In the crystal structure, the side chains of these residues, which are located in helix A, the AB loop, helix B, and helix D, are surface accessible and are clustered in space, thus constituting an epitope for CNTFR alpha. By the same analysis, a partial epitope for gp130 was also identified on the surface of helix A that faces away from the alpha-epitope. Superposition of the CNTF and growth hormone structures showed that the location of these epitopes on CNTF is analogous to the location of the first and second receptor epitopes on the surface of growth hormone. Further comparison with proposed binding sites for alpha- and beta-receptors on interleukin-6 and leukemia inhibitory factor indicated that this epitope topology is conserved among helical cytokines. In each case, epitope I is utilized by the specificity-conferring component, whereas epitopes II and III are used by accessory components. Thus, in addition to a common fold, helical cytokines share a conserved order of receptor epitopes that is function related.
Panayotatos
N
N
REGENERON Pharmaceuticals Inc., Tarrytown, New York 10591-6707, USA.
Radziejewska
E
E
Acheson
A
A
Somogyi
R
R
Thadani
A
A
Hendrickson
W A
WA
McDonald
N Q
NQ
D000818:Animals; D001665:Binding Sites; D002645:Chickens; D020934:Ciliary Neurotrophic Factor; D016207:Cytokines / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D000939:Epitopes; D013006:Growth Hormone / Q000378:metabolism; D006131:Growth Inhibitors / Q000378:metabolism; D006801:Humans; D015850:Interleukin-6 / Q000378:metabolism; D053605:Leukemia Inhibitory Factor; D008222:Lymphokines / Q000378:metabolism; D009419:Nerve Tissue Proteins / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D017433:Protein Structure, Secondary; D020801:Receptor, Ciliary Neurotrophic Factor; D017475:Receptors, Nerve Growth Factor / Q000276:immunology* / Q000378:metabolism
D020934:Ciliary Neurotrophic Factor; D016207:Cytokines; D000939:Epitopes; D006131:Growth Inhibitors; D015850:Interleukin-6; C507837:LIF protein, human; D053605:Leukemia Inhibitory Factor; D008222:Lymphokines; D009419:Nerve Tissue Proteins; D020801:Receptor, Ciliary Neurotrophic Factor; D017475:Receptors, Nerve Growth Factor; D013006:Growth Hormone