title: Spatial orientation of the alpha and betac receptor chain binding sites on monomeric human interleukin-5 constructs.
abstract: Interleukin-5 (IL-5), a disulfide-linked homodimer, can be induced to fold as a biological active monomer by extending the loop between its third and fourth helices (Dickason, R. R., and Huston, D. P. (1996) Nature 379, 652-655). We have designed eight monomeric IL-5 proteins to optimize biological activity and stability of the monomer. This was achieved by (i) inserting the joining loop at three different positions, (ii) by introducing an additional intramolecular disulfide bridge onto these backbones, and (iii) by creating circular permutations to fix the position of the carboxyl-terminal helix relative to the three other helices. The proteins dimerize with Kd values ranging from 20 to 200 microM and are therefore monomeric at the picomolar concentrations where they are biologically active. Introduction of a second disulfide confers increased stability, but this increased rigidity results in lower activity of the protein. Contrary to wild type IL-5, mutation of the betac contact residue on the first helix, Glu12, to Lys, into the circularly permutated constructs, did not abolish TF-1 proliferative and eosinophil activation activities. These results indicate that activation of the IL-5 receptor complex is not mediated solely by Glu12 on the first helix, and alternative mechanisms are discussed.
authors.affiliation: Geneva Biomedical Research Institute, 1228 Plan-les-Ouates, Geneva, Switzerland.
authors.forename: M D
authors.initials: MD
authors.lastname: Edgerton
authors.forename: P
authors.initials: P
authors.lastname: Graber
authors.forename: D
authors.initials: D
authors.lastname: Willard
authors.forename: T
authors.initials: T
authors.lastname: Consler
authors.forename: M
authors.initials: M
authors.lastname: McKinnon
authors.forename: I
authors.initials: I
authors.lastname: Uings
authors.forename: C Y
authors.initials: CY
authors.lastname: Arod
authors.forename: F
authors.initials: F
authors.lastname: Borlat
authors.forename: R
authors.initials: R
authors.lastname: Fish
authors.forename: M C
authors.initials: MC
authors.lastname: Peitsch
authors.forename: T N
authors.initials: TN
authors.lastname: Wells
authors.forename: A E
authors.initials: AE
authors.lastname: Proudfoot
mesh_terms: D001665:Binding Sites; D019281:Dimerization; D006801:Humans; D015848:Interleukin-5 / Q000737:chemistry*; D017433:Protein Structure, Secondary; D018123:Receptors, Interleukin / Q000737:chemistry*; D053649:Receptors, Interleukin-5
chemical_list: D015848:Interleukin-5; D018123:Receptors, Interleukin; D053649:Receptors, Interleukin-5