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The Influence of Adnectin Binding on the Extracellular Domain of Epidermal Growth Factor Receptor

  • Research Article
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Journal of The American Society for Mass Spectrometry

Abstract

The precise and unambiguous elucidation and characterization of interactions between a high affinity recognition entity and its cognate protein provides important insights for the design and development of drugs with optimized properties and efficacy. In oncology, one important target protein has been shown to be the epidermal growth factor receptor (EGFR) through the development of therapeutic anticancer antibodies that are selective inhibitors of EGFR activity. More recently, smaller protein derived from the 10th type III domain of human fibronectin termed an adnectin has also been shown to inhibit EGFR in clinical studies. The mechanism of EGFR inhibition by either an adnectin or an antibody results from specific binding of the high affinity protein to the extracellular portion of EGFR (exEGFR) in a manner that prevents phosphorylation of the intracellular kinase domain of the receptor and thereby blocks intracellular signaling. Here, the structural changes induced upon binding were studied by probing the solution conformations of full length exEGFR alone and bound to a cognate adnectin through hydrogen/deuterium exchange mass spectrometry (HDX MS). The effects of binding in solution were identified and compared with the structure of a bound complex determined by X-ray crystallography.

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Acknowledgments

The authors thank Professor Thomas E. Wales for helpful discussions. The authors also thank Dr. Bruce Car, Dr. Morrey Atkinson, and Dr. Peter Moesta from Bristol-Myers Squibb Company for their support of this project. This work was supported in part by grants from the National Institute of Health (GM086507 and GM 101135) and a research collaboration with the Waters Corporation.

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  1. Joomi Ahn

    Present address: Biological Sciences, Allergan, Inc., 2525 Dupont Drive, Irvine, CA, 92612, USA

Authors and Affiliations

  1. Department of Chemistry and Chemical Biology, Northeastern University, Boston, MA, 02115, USA

    Roxana E. Iacob & John R. Engen

  2. Bioanalytical and Discovery Analytical Sciences, Bristol-Myers Squibb Company, Hopewell, NJ, 08534, USA

    Guodong Chen, Hui Wei, Jingjie Mo & Adrienne A. Tymiak

  3. Waters Corporation, Milford, MA, 01757, USA

    Joomi Ahn & Stephane Houel

  4. Biologics Manufacturing and Process Development, Global Manufacturing and Supply, Bristol-Myers Squibb Company, Hopewell, NJ, 08534, USA

    Li Tao

  5. Protein Science, Research and Development, Bristol-Myers Squibb Company, Princeton, NJ, 08534, USA

    Daniel Cohen, Dianlin Xie, Zheng Lin, Paul E. Morin & Michael L. Doyle

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  1. Roxana E. Iacob
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  2. Guodong Chen
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Iacob, R.E., Chen, G., Ahn, J. et al. The Influence of Adnectin Binding on the Extracellular Domain of Epidermal Growth Factor Receptor. J. Am. Soc. Mass Spectrom. 25, 2093–2102 (2014). https://doi.org/10.1007/s13361-014-0973-1

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