Evidence that natural murine soluble interleukin 4 receptors may act as transport proteins

Rafael Fernandez-Botran, Ellen S. Vitetta

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Abstract

The present studies were undertaken to determine whether the interleukin 4 binding proteins (IL-4BPs) previously identified in the biological fluids of mice are soluble forms of IL-4Rs. We also studied the binding properties of IL-4BPs in order to gain insight into their physiological role in vivo. Affinity-purified IL-4BPs and recombinant soluble IL-4Rs generated similar one-dimensional (Cleveland) peptide maps after digestion with either Staphylococcus aureus V8 protease or trypsin, indicating structural similarities. Furthermore, a rat mAb directed against the murine IL-4Rs immunoprecipitated the IL-4BPs and completely inhibited binding of 125I-IL-4 to a purified preparation of IL-4BPs. Taken together these data indicate that the IL-4BPs are soluble IL-4Rs. At 4°C the IL-4BPs competitively inhibited the binding of IL-4 to membrane IL-4Rs but their ability to prevent binding of IL-4 to cells at 37°C, at the same concentrations, was significantly reduced. Kinetic binding studies of soluble IL-4BPs vs. membrane IL-4Rs disclosed important differences in their rates of dissociation from IL-4. Whereas dissociation at 4°C was slow for both, dissociation of IL-4 from IL-BPs at 37°C was considerably faster (t1/2 of 2 min) than dissociation of IL-4 from membrane IL-4Rs (t1/2 of ∼69 min). Temperature-dependent changes in dissociation kinetics were reversible, and could not be accounted for by either inactivation of the IL-4BPs at 37°C or receptor internalization. Additional experiments also demonstrated that when IL-4BPs bind to IL-4 at 37°C, the IL-4/IL-4BPs complex can rapidly dissociate, allowing IL-4 to bind to membrane IL-4Rs. In addition, binding of IL-4 by the IL-4BPs protects IL-4 from proteolytic degradation. Taken together, these results suggest that the IL-4BPs are naturally occurring forms of soluble IL-4Rs and that some of their properties (fast dissociation kinetics and protection of IL-4 from proteolysis) are consistent with a potential role as carrier proteins for IL-4 in the circulation.

Original languageEnglish (US)
Pages (from-to)673-681
Number of pages9
JournalJournal of Experimental Medicine
Volume174
Issue number3
StatePublished - 1991

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Interleukin-4 Receptors
Interleukin-4
Carrier Proteins
Membranes

ASJC Scopus subject areas

  • Immunology

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Evidence that natural murine soluble interleukin 4 receptors may act as transport proteins. / Fernandez-Botran, Rafael; Vitetta, Ellen S.

In: Journal of Experimental Medicine, Vol. 174, No. 3, 1991, p. 673-681.

Research output: Contribution to journalArticle

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