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Interleukin-3 Binding to the Murine β IL-3 and Human βc Receptors Involves Functional Epitopes Formed by Domains 1 and 4 of Different Protein Chains

Murphy, James; Ford, Sally; Olsen, Jane; Gustin, Sonya; Jeffrey, Peter; Ollis, David; Young, Ian

Description

Interleukin-3 (IL-3) is a cytokine produced by activated T-cells and mast cells that is active on a broad range of hematopoietic cells and in the nervous system and appears to be important in several chronic inflammatory diseases. In this study, alanine substitutions were used to investigate the role of residues of the human β-common (hβc) receptor and the murine IL-3-specific (βIL-3) receptor in IL-3 binding. We show that the domain 1 residues, Tyr15 and Phe79, of the hβc receptor are...[Show more]

dc.contributor.authorMurphy, James
dc.contributor.authorFord, Sally
dc.contributor.authorOlsen, Jane
dc.contributor.authorGustin, Sonya
dc.contributor.authorJeffrey, Peter
dc.contributor.authorOllis, David
dc.contributor.authorYoung, Ian
dc.date.accessioned2015-12-13T22:41:11Z
dc.date.available2015-12-13T22:41:11Z
dc.identifier.issn0021-9258
dc.identifier.urihttp://hdl.handle.net/1885/78398
dc.description.abstractInterleukin-3 (IL-3) is a cytokine produced by activated T-cells and mast cells that is active on a broad range of hematopoietic cells and in the nervous system and appears to be important in several chronic inflammatory diseases. In this study, alanine substitutions were used to investigate the role of residues of the human β-common (hβc) receptor and the murine IL-3-specific (βIL-3) receptor in IL-3 binding. We show that the domain 1 residues, Tyr15 and Phe79, of the hβc receptor are important for high affinity IL-3 binding and receptor activation as shown previously for the related cytokines, interleukin-5 and granulocyte-macrophage colony-stimulating factor, which also signal through this receptor subunit. From the x-ray structure of hβc, it is clear that the domain 1 residues cooperate with domain 4 residues to form a novel ligand-binding interface involving the two protein chains of the intertwined homodimer receptor. We demonstrate by ultracentrifugation that the βIL-3 receptor is also a homodimer. Its high sequence homology with hβc suggests that their structures are homologous, and we identified an analogous binding interface in βIL-3 for direct IL-3 binding to the high affinity binding site in hβc. Tyr21 (A-B loop), Phe85, and Asn87 (E-F loop) of domain 1; Ile320 of the interdomain loop; and Tyr348 (B′-C′ loop) and Tyr401 (F′-G′ loop) of domain 4 were shown to have critical individual roles and Arg84 and Tyr317 major secondary roles in direct murine IL-3 binding to the βIL-3 receptor. Most surprising, none of the key residues for direct IL-3 binding were critical for high affinity binding in the presence of the murine IL-3 α receptor, indicating a fundamentally different mechanism of high affinity binding to that used by hβc.
dc.publisherAmerican Society for Biochemistry and Molecular Biology Inc
dc.sourceJournal of Biological Chemistry
dc.subjectKeywords: Activation analysis; Cells; Disease control; Neurology; Signal processing; X ray analysis; Hematopoietic cells; Inflamatory diseases; T-cells; Binding energy; asparagine; epitope; granulocyte macrophage colony stimulating factor; interleukin 3; interleuki
dc.titleInterleukin-3 Binding to the Murine β IL-3 and Human βc Receptors Involves Functional Epitopes Formed by Domains 1 and 4 of Different Protein Chains
dc.typeJournal article
local.description.notesImported from ARIES
local.description.refereedYes
local.identifier.citationvolume279
dc.date.issued2004
local.identifier.absfor030499 - Medicinal and Biomolecular Chemistry not elsewhere classified
local.identifier.ariespublicationMigratedxPub7053
local.type.statusPublished Version
local.contributor.affiliationMurphy, James, College of Medicine, Biology and Environment, ANU
local.contributor.affiliationFord, Sally, College of Medicine, Biology and Environment, ANU
local.contributor.affiliationOlsen, Jane, College of Medicine, Biology and Environment, ANU
local.contributor.affiliationGustin, Sonya, College of Medicine, Biology and Environment, ANU
local.contributor.affiliationJeffrey, Peter, College of Medicine, Biology and Environment, ANU
local.contributor.affiliationOllis, David, College of Physical and Mathematical Sciences, ANU
local.contributor.affiliationYoung, Ian, College of Medicine, Biology and Environment, ANU
local.bibliographicCitation.issue25
local.bibliographicCitation.startpage26500
local.bibliographicCitation.lastpage26508
local.identifier.doi10.1074/jbc.M402705200
dc.date.updated2015-12-11T10:00:13Z
local.identifier.scopusID2-s2.0-2942729678
CollectionsANU Research Publications

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