Molecular basis of the membrane-anchored and two soluble isoforms of the human interleukin 5 receptor alpha subunit.

Tavernier, Jan; Tuypens, T; Plaetinck, Geert; Verhee, Annick; Fiers, Walter; Devos, René

doi:10.1073/pnas.89.15.7041

Cited by 124 publications

(91 citation statements)

References 27 publications

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“…Two major nonexclusive mechanisms are responsible for the generation of soluble cytokine receptors: proteolytic cleavage of transmembrane receptors catalyzed mostly by metalloproteases [15][16][17][18][19] or de novo synthesis of alternatively spliced mRNAs encoding soluble receptor molecules lacking a transmembrane domain. [20][21][22][23][24][25][26][27][28] Soluble cytokine receptors appear to be potent regulators of cytokine activities. Interference with the binding of cytokines to their membrane receptors and, thus, inhibition of cytokine signaling has been demonstrated to occur in vitro and in vivo.…”

Section: Introductionmentioning

confidence: 99%

A soluble form of the murine common γ chain is present at high concentrations in vivo and suppresses cytokine signaling

Meißner

Blum

Schnare

et al. 2001

Blood

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The common gamma-chain (␥c) is a component of the receptors for IL-2, IL-4, IL-7, IL-9, and IL-15 and is essential for their signal transduction. Western blotting and a newly established enzyme-linked immunosorbent assay detected substantial constitutive levels (50-250 ng/mL) of soluble ␥c (s␥c) in sera of murine inbred strains. It was demonstrated that purified immune cells, such as T, B, and natural killer cells, and macrophages released this protein after activation. Transfection experiments with cDNA encoding the fulllength ␥c showed that shedding of the transmembrane receptor led to the release of s␥c. The shedding enzymes, however, appeared to be distinct from those cleaving other cytokine receptors because inhibitors of metalloproteases (eg, TAPI) did not influence s␥c release. In vivo, superantigen-induced stimulation of T cells enhanced s␥c serum concentrations up to 10-fold within 6 hours. Because these findings demonstrated regulated expression of a yet unknown molecule in the immune response, further experiments were performed to assess the possible function(s) of s␥c. A physiological role of s␥c was indicated by its capacity to specifically inhibit cell growth induced by ␥c-dependent cytokines. Mutational analysis revealed that the Cterminus and the WSKWS motif are essential for the cytokine inhibitory effect of the s␥c and for binding of the molecule to cytokine receptor-expressing cells. Thus, competitive displacement of the transmembrane ␥c by excess s␥c is the most likely mechanism of cell growth inhibition. It was implied that naturally produced s␥c is a negative modulator of ␥c-dependent cytokines. IntroductionThe murine common gamma-chain (␥c) is a 64-kd type 1 transmembrane protein of the cytokine receptor family. 1 Although the ␥c alone is unable to bind to cytokines, it has been shown to be an essential component of the cell surface receptor complexes of IL-2, IL-4, IL-7, IL-9, and IL-15. 1 Interaction of the respective cytokines with the ␥c within the receptor complexes results in the activation of the Janus kinase JAK3, 2 which subsequently interacts with Pyk2, a member of focal adhesion tyrosine kinases, leading to the phosphorylation of this protein. 3 Recent studies have shown that the ␥c is critical for lymphoid development because genetic defects of either this molecule 4 or JAK3 5 in humans or targeted deletions of the ␥c 6,7 or JAK3 8-10 in mice severely impair the development of T and B cells, leading to severe combined immune deficiency syndromes (SCID). 11 Soluble cytokine receptors and growth factor receptors are present as immunomodulatory molecules in body fluids of humans and mice (reviewed in references 12 and 14). Two major nonexclusive mechanisms are responsible for the generation of soluble cytokine receptors: proteolytic cleavage of transmembrane receptors catalyzed mostly by metalloproteases [15][16][17][18][19] or de novo synthesis of alternatively spliced mRNAs encoding soluble receptor molecules lacking a transmembrane domain. [20][21][22][23][24][25][26][27][28] ...

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Section: Introductionmentioning

confidence: 99%

A soluble form of the murine common γ chain is present at high concentrations in vivo and suppresses cytokine signaling

Meißner

Blum

Schnare

et al. 2001

Blood

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“…The a chain of the human IL-5 receptor binds human IL-5 with an affinity of about 2 nM, the binding affinity of the heterodimeric native IL-5 receptor consisting of the a and /l chain is only 2-4-fold higher [4,11, 121. Thus synthetic compounds that inhibit the interaction between IL-5 and the IL-5 -receptor a chain can be expected io be as effective in inhibiting binding of IL-5 to its heterodimeric native receptor and in preventing IL-5 -mediated cell signaling.…”

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confidence: 99%

“…Similarly to the a chain of the IL-5 receptor, the a chains of the GM-CSF and IL-3 receptors bind specifically their cognate ligand and associate with the common p chain shared by all three receptors for signal transduction [4, 91. While the / 3 chain is required for signal transduction, some specificity for the transduced signal appears to be related to the a chain since divergent signaling pathways appear to be induced by either the GM-CSF and IL-3 receptors or the receptor for IL-5 [lo].The a chain of the human IL-5 receptor binds human IL-5 with an affinity of about 2 nM, the binding affinity of the heterodimeric native IL-5 receptor consisting of the a and /l chain is only 2-4-fold higher [4,11, 121. Thus synthetic compounds that inhibit the interaction between IL-5 and the IL-5 -receptor a chain can be expected io be as effective in inhibiting binding of IL-5 to its heterodimeric native receptor and in preventing IL-5 -mediated cell signaling.…”

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confidence: 99%

Characterization of Potential Antagonists of Human Interleukin 5 Demonstrates Their Cross‐Reactivity with Receptors for Interleukin 3 and Granulocyte‐Macrophage Colony‐Stimulating Factor

Wiekowski¹,

Prosser²,

Taremi³

et al. 1997

European Journal of Biochemistry

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The ligand-binding a-chain of the human interleukin 5 (IL-5) receptor was expressed in its soluble form, lacking the transmembrane and cytoplasmic domains, from recombinant baculovirus. The soluble receptor was used in a scintillation proximity assay to identify two chemical compounds that inhibit binding of human IL-5 to the soluble receptor a chain with IC,, of 8 pM and 11 pM, These compounds also inhibited the interaction of human IL-5 with its membrane-bound receptor, composed of the ligandbinding a chain and signal-transducing p chain, and prevented signaling through the receptor. Analysis by surface plasmon resonance and matrix-assisted laser-desorptiodionization mass spectrometry showed that the identified compounds bound irreversibly to the receptor at a 1 : 1 (mol/mol) ratio, suggesting a covalent interaction with the a chain of the human IL-5 receptor. Both compounds also inhibited the interaction of the receptors for interleukin 3 (IL-3) and granulocyte-macrophage colony-stimulating factor (GM-CSF), which are involved in hematopoietic differentiation and activation of immune cells, thus eliminating them as potential therapeutic agents. The inhibition of the strucuturally closely related receptors for IL-5, IL-3 and GM-CSF by both compounds, while binding of interleukin-4 to its receptor was not affected, suggests that a similar reactive site exists in the ligand-binding domains of the receptors for IL-5, IL-3 and GM-CSF. Keywords : human interleukin 5 ; interleukin-5 receptor; interleukin-3 receptor; granulocyte-macrophagecolony-stimulating-factor receptor; receptor antagonist.Human interleukin (IL-5) is a disulfide-linked homodimeric cytokine produced from T cells that promotes proliferation and differentiation of eosinophils [l, 21. The observed effect of IL-5 on eosinophils suggests that IL-5 has a role in the pathogenic inflammatory responses of hypersensitivity and other diseases in which eosinophil infiltration has been observed. Antagonists of IL-5 might serve as drugs for the treatment of allergic diseases such as bronchial asthma and other hypereosinophilic syndromes.Human IL-5 transduces its signal by binding to its specific receptor expressed on eosinophils [3]. The human IL-5 receptor has been identified as a heterodimer consisting of the ligandbinding a chain of 60 kDa and a signal-transducing /l chain of 120 kDa [4]. Association of the a chain with the p chain is required for the induction of signal-transduction pathways [4, 51 involving Jak2 and STAT1 (signal transducer and activator of transcription 1) or Lyn and the mitogen-activated protein kinases [6-81.The IL-5 receptor and the receptors for interleukin 3 (IL-3) and granulocyte-macrophage colony-stimulating factor (GM- CSF) form a subfamily of cytokine receptors based on shared structural and functional features. Similarly to the a chain of the IL-5 receptor, the a chains of the GM-CSF and IL-3 receptors bind specifically their cognate ligand and associate with the common p chain shared by all three receptors for signal transductio...

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“…To enhance the solubility and binding signal of the peptide in surface plasmon resonance (SPR) assays, we employed a recombinant system to fuse the peptide to a highly soluble protein, thioredoxin. We used thioredoxin-fused AF17121 for kinetic interaction analysis with soluble IL5R␣ (sIL5R␣; extracellular domain of IL5R␣) (22) as well as for biological inhibition assays. Alanine substitution of the two half-cystine residues in AF17121 completely impaired its binding to IL5R␣ and its corresponding inhibitory action on cells.…”

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confidence: 99%

Receptor Epitope Usage by an Interleukin-5 Mimetic Peptide

Ishino

Urbina

Bhattacharya

et al. 2005

Journal of Biological Chemistry

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The cyclic peptide AF17121 is a library-derived antagonist for human interleukin-5 (IL5) receptor ␣ (IL5R␣) and inhibits IL5 activity. Our previous results have demonstrated that the sixth arginine residue of the peptide is crucial for the inhibitory effect and that several acidic residues in the N-and C-terminal regions also make a contribution, although to a lesser extent (Ruchala, P., Varadi, G., Ishino, T., Scibek, J., Bhattacharya, M., Urbina, C., Van Ryk, D., Uings, I., and Chaiken, I. (2004) Biopolymers 73, 556 -568). However, the recognition mechanism of the receptor has remained unresolved. In this study, AF17121 was fused to thioredoxin by recombinant DNA techniques and examined for IL5R␣ interaction using a surface plasmon resonance biosensor method. Kinetic analysis revealed that the dissociation rate of the peptide⅐receptor complex is comparable with that of the cytokine⅐receptor complex. The fusion peptide competed with IL5 for both biological function and interaction with IL5R␣, indicating that the binding sites on the receptor are shared by AF17121 and IL5. To define the epitope residues for AF17121, we defined its Asthma is an incurable disease characterized by eosinophil bronchial inflammation and tissue remodeling of the airway wall (1). Human interleukin (IL) 1 -5 is a key cytokine that plays a critical role in the differentiation, proliferation, migration, and activation of eosinophils and has been implicated in the pathogenesis of eosinophil-associated allergic inflammation such as asthma (2, 3). Injection of IL5 increases eosinophil numbers in the blood, bone marrow, spleen, and peritoneal cavities (4, 5). This increase can be prevented by the passive administration of anti-IL5 or anti-IL5 receptor antibodies (5, 6). One distinguishing characteristic of IL5 versus other cytokines is that IL5 functions selectively to activate eosinophils (7). These data argue that IL5 plays a central role in the development of allergen-induced eosinophils and suggest that the ability to block IL5 activity evokes the potential for alternative treatment for asthma.At the molecular level, IL5 leads to biological functions through recruitment of a cell-surface receptor composed of two polypeptide chains, ␣ and ␤ (8). The ␣ chain is IL5-specific and is called IL5 receptor ␣ (IL5R␣), whereas the ␤ chain is shared with IL3 and granulocyte/macrophage colony-stimulating factor (GM-CSF) (9 -11) and is called the common ␤ chain (␤c). Despite a high degree of amino acid sequence similarity of the ␣ chains for IL5, IL3, and GM-CSF, their interaction with cognate cytokine is strictly specific, and no cross-reactivity is found among these cytokines. Previous observations argue that receptor subunit recruitment occurs stepwise, with initial formation of the IL5⅐IL5R␣ complex required for ␤c binding to induce cytoplasmic signal transduction (12). IL5R␣ alone binds IL5 with an equilibrium dissociation constant of 0.8 nM when expressed in COS cells, and this binding affinity is increased by only 1.5-fold when ␣ and ␤ chains a...

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Molecular basis of the membrane-anchored and two soluble isoforms of the human interleukin 5 receptor alpha subunit.

Cited by 124 publications

References 27 publications

A soluble form of the murine common γ chain is present at high concentrations in vivo and suppresses cytokine signaling

A soluble form of the murine common γ chain is present at high concentrations in vivo and suppresses cytokine signaling

Characterization of Potential Antagonists of Human Interleukin 5 Demonstrates Their Cross‐Reactivity with Receptors for Interleukin 3 and Granulocyte‐Macrophage Colony‐Stimulating Factor

Receptor Epitope Usage by an Interleukin-5 Mimetic Peptide

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