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Asymmetric usage of antagonist charged residues drives interleukin-5 receptor recruitment but is insufficient for receptor activation
Journal article   Open access   Peer reviewed

Asymmetric usage of antagonist charged residues drives interleukin-5 receptor recruitment but is insufficient for receptor activation

Tetsuya Ishino, Udaya Pillalamarri, Dominick Panarello, Madhushree Bhattacharya, Cecilia Urbina, Stephanie Horvat, Sanjay Sarkhel, Bradford Jameson and Irwin Chaiken
Biochemistry (Easton), v 45(4), pp 1106-1115
31 Jan 2006
DOI: https://doi.org/10.1021/bi0518038
PMID: 16430207
Featured in Collection :   UN Sustainable Development Goals @ Drexel
url
https://doi.org/10.1021/bi0518038View
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Abstract

Surface Plasmon Resonance Temperature Epitopes - metabolism Receptors, Interleukin - chemistry Humans Molecular Sequence Data Interleukin-5 Receptor alpha Subunit Peptides, Cyclic - pharmacology Structure-Activity Relationship Histidine - metabolism Recombinant Fusion Proteins - metabolism Peptides, Cyclic - chemistry Peptides, Cyclic - metabolism Receptors, Interleukin - metabolism Thioredoxins - genetics Thioredoxins - metabolism Protein Structure, Tertiary Amino Acid Sequence Models, Molecular Receptors, Interleukin - antagonists & inhibitors Arginine - chemistry Animals Protein Binding Receptors, Interleukin-5 Recombinant Fusion Proteins - genetics Epitopes - chemistry Drosophila - metabolism Kinetics Mutation Histidine - chemistry Arginine - metabolism
The cyclic peptide AF17121 (VDECWRIIASHTWFCAEE) is a library-derived antagonist for human Interleukin-5 receptor alpha (IL5Ralpha). We have previously demonstrated that AF17121 mimics Interleukin-5 (IL5) by binding in a region of IL5Ralpha that overlaps the IL5 binding epitope. In the present study, to explore the functional importance of the amino acid residues of AF17121 required for effective binding to, and antagonism of, IL5Ralpha, each charged residue was subjected to site-directed mutagenesis and examined for IL5Ralpha interaction by using a surface plasmon resonance biosensor. One residue, Arg(6), was found to be essential for receptor antagonism; its replacement with either alanine or lysine completely abolished the interaction between AF17121 and IL5Ralpha. Other charged residues play modulatory roles. One class consists of the N-terminal acidic cluster (Asp(2) and Glu(3)) for which alanine replacement decreased the association rate. A second class consists of His(11) and the C-terminal acidic cluster (Glu(17) and Glu(18)) for which alanine replacement increased the dissociation rate. Binding model analysis of the mutants of the latter class of residues indicated the existence of conformational rearrangement during the interaction. On the basis of these results, we propose a model in which Arg(6) and N-terminal acidic residues drive the encounter complex, while Arg(6), His(11), and C-terminal acidic residues are involved in stabilizing the final complex. These data argue that the charged residues of AF17121 are utilized asymmetrically in the pathway of inhibitor-receptor complex formation to deactivate the receptor function. The results also help focus emerging models for the mechanism by which IL5 activates the IL5Ralpha-betac receptor system.

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Web of Science research areas
Biochemistry & Molecular Biology
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