This article is a US Government work, and, as such, is in the public domain in the United States of America.
Mapping mouse IL-13 binding regions using structure modeling, molecular docking, and high-density peptide microarray analysis†
Article first published online: 9 NOV 2010
Published 2010 Wiley-Liss, Inc.
Proteins: Structure, Function, and Bioinformatics
Volume 79, Issue 1, pages 282–293, January 2011
How to Cite
Madala, S. K., Dolan, M. A., Sharma, D., Ramalingam, T. R., Wilson, M. S., Mentink-Kane, M. M., Masison, D. C. and Wynn, T. A. (2011), Mapping mouse IL-13 binding regions using structure modeling, molecular docking, and high-density peptide microarray analysis . Proteins, 79: 282–293. doi: 10.1002/prot.22881
- Issue published online: 30 NOV 2010
- Article first published online: 9 NOV 2010
- Accepted manuscript online: 22 SEP 2010 02:29PM EST
- Manuscript Accepted: 27 AUG 2010
- Manuscript Revised: 26 JUL 2010
- Manuscript Received: 10 MAY 2010
- Intramural Research Program of the NIH, NIAID
- cytokine receptors;
- peptide arrays;
- protein-protein interactions;
- structure and protein modeling
Interleukin-13 is a Th2-associated cytokine responsible for many pathological responses in allergic asthma including mucus production, inflammation, and extracellular matrix remodeling. In addition, IL-13 is required for immunity to many helminth infections. IL-13 signals via the type-II IL-4 receptor, a heterodimeric receptor of IL-13Rα1 and IL-4Rα, which is also used by IL-4. IL-13 also binds to IL-13Rα2, but with much higher affinity than the type-II IL-4 receptor. Binding of IL-13 to IL-13Rα2 has been shown to attenuate IL-13 signaling through the type-II IL-4 receptor. However, molecular determinants that dictate the specificity and affinity of mouse IL-13 for the different receptors are largely unknown. Here, we used high-density overlapping peptide arrays, structural modeling, and molecular docking methods to map IL-13 binding sequences on its receptors. Predicted binding sequences on mouse IL-13Rα1 and IL-13Rα2 were in agreement with the reported human IL-13 receptor complex structures and site-directed mutational analysis. Novel structural differences were identified between IL-13 receptors, particularly at the IL-13 binding interface. Notably, additional binding sites were observed for IL-13 on IL-13Rα2. In addition, the identification of peptide sequences that are unique to IL-13Rα1 allowed us to generate a monoclonal antibody that selectively binds IL-13Rα1. Thus, high-density peptide arrays combined with molecular docking studies provide a novel, rapid, and reliable method to map cytokine-receptor interactions that may be used to generate signaling and decoy receptor-specific antagonists. Proteins 2010. © 2010 Wiley-Liss, Inc.