Molecular basis of a high affinity murine interleukin-5 receptor.
The mouse interleukin-5 receptor (mIL-5R) consists of two components one of which, the mIL-5R alpha-chain, binds mIL-5 with low affinity. Recently we demonstrated that monoclonal antibodies (Mabs) recognizing the second mIL-5R beta-chain, immunoprecipitate a p130-140 protein doublet which corresponds to the mIL-3R and the mIL-3R-like protein, the latter chain for which so far no ligand has been identified. In this study we show that a high affinity mIL-5R can be reconstituted on COS1 cells by co-expression of the mIL-5R alpha-chain with the mIL-3R-like protein (beta-chain). Cross-linking of 125I-labeled mIL-5 to the COS1 cells co-transfected with both cDNAs revealed the same pattern as in B13 cells, i.e. two proteins of 60 and 130 kd which correspond to the low affinity mIL-5R alpha-chain and the mIL-3R-like protein, respectively. The dissociation rate of mIL-5 from this reconstituted high affinity site was lower than that of the low affinity site, whereas the association rate was unchanged. Nonetheless, the apparent dissociation constant (Kd) for this reconstituted receptor was still 10-fold higher than the Kd observed for B13 cells. Although the mIL-3R is greater than 90% homologous to the mIL-3R-like protein, no increase in affinity for mIL-5 was detected on COS1 cells co-transfected with the cDNAs for the mIL-5R alpha-chain and the mIL-3R protein.
Devos
R
R
Roche Research Gent, Belgium.
Plaetinck
G
G
Van der Heyden
J
J
Cornelis
S
S
Vandekerckhove
J
J
Fiers
W
W
Tavernier
J
J
D000818:Animals; D000911:Antibodies, Monoclonal; D001483:Base Sequence; D002460:Cell Line; D003432:Cross-Linking Reagents; D004247:DNA / Q000235:genetics; D015848:Interleukin-5 / Q000378:metabolism*; D007700:Kinetics; D051379:Mice; D008969:Molecular Sequence Data; D011233:Precipitin Tests; D011971:Receptors, Immunologic / Q000235:genetics* / Q000276:immunology; D018123:Receptors, Interleukin; D053649:Receptors, Interleukin-5; D013379:Substrate Specificity; D014162:Transfection
D000911:Antibodies, Monoclonal; D003432:Cross-Linking Reagents; D015848:Interleukin-5; D011971:Receptors, Immunologic; D018123:Receptors, Interleukin; D053649:Receptors, Interleukin-5; D004247:DNA