Linked Life Data

8892059

Source:http://linkedlifedata.com/resource/pubmed/id/8892059

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
9
pubmed:dateCreated
1997-1-29
pubmed:abstractText
Interleukin (IL) 5 specifically induces the differentiation of eosinophils which are central to the pathogenesis of allergies and asthma. Structurally, IL-5 is a unique member of the short-chain helical bundle subfamily of cytokines. In contrast to other subfamily members which fold unimolecularly into a single helical bundle, IL-5 forms a pair of helical bundles by the interdigitation of two identical monomers covalently linked by a pair of intermolecular disulfide bonds. Although a native IL-5 monomer lacks bioactivity, we recently reported the engineering of an insertional mutant of IL-5 (designated mono5) which folds unimolecularly into a single helical bundle and has biological activity similar to that of native IL-5. Here we demonstrate no differences in signal transduction pathways utilized by mono5 and IL-5, as determined by western blot analysis of early tyrosine phosphorylation events, Jak2 activation, and mitogen-activated protein kinase activation. However, binding studies utilizing conformationally dependent neutralizing anti-IL-5 monoclonal antibodies localized a tertiary structural perturbation near the insert of mono5. This perturbation enabled localization of a limited region of the tertiary structure of IL-5 that engages the IL-5 receptor alpha-chain. Fluorescent labeling studies further revealed that the cysteines of mono5 contained free sulfhydryl groups, thereby demonstrating that the role of the disulfide bonds of IL-5 is the structural maintenance of other functional domains. The retention of conformation epitopes by mono5, but not IL-5, under reducing conditions and the equivalent thermostability of mono5 and IL-5 despite the absence of a disulfide bond in mono5 indicated that the conformation assumed by mono5 is very stable. In addition to providing the structural framework for designing novel IL-5 agonists and antagonists, the knowledge gained from the development of mono5 will enable other helical bundle proteins to be redesigned with therapeutic potential.
pubmed:grant
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Sep
pubmed:issn
0946-2716
pubmed:author
pubmed:issnType
Print
pubmed:volume
74
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
535-46
pubmed:dateRevised
2011-7-8
pubmed:meshHeading
pubmed-meshheading:8892059-Antibodies, Monoclonal, pubmed-meshheading:8892059-Blotting, Western, pubmed-meshheading:8892059-Chromatography, High Pressure Liquid, pubmed-meshheading:8892059-Epitopes, pubmed-meshheading:8892059-Hypersensitivity, pubmed-meshheading:8892059-Inflammation, pubmed-meshheading:8892059-Interleukin-5, pubmed-meshheading:8892059-Janus Kinase 1, pubmed-meshheading:8892059-Models, Molecular, pubmed-meshheading:8892059-Phosphotyrosine, pubmed-meshheading:8892059-Protein Binding, pubmed-meshheading:8892059-Protein Conformation, pubmed-meshheading:8892059-Protein Engineering, pubmed-meshheading:8892059-Protein Structure, Tertiary, pubmed-meshheading:8892059-Protein-Tyrosine Kinases, pubmed-meshheading:8892059-Signal Transduction, pubmed-meshheading:8892059-Sulfhydryl Compounds, pubmed-meshheading:8892059-Sulfhydryl Reagents, pubmed-meshheading:8892059-Temperature, pubmed-meshheading:8892059-Tumor Cells, Cultured
pubmed:year
1996
pubmed:articleTitle
Engineering of a functional interleukin-5 monomer: a paradigm for redesigning helical bundle cytokines with therapeutic potential in allergy and asthma.
pubmed:affiliation
Baylor College of Medicine, Department of Microbiology and Immunology, Houston, TX 77030, USA.
pubmed:publicationType
Journal Article, Research Support, U.S. Gov't, P.H.S., Research Support, Non-U.S. Gov't