Synthesis and biochemical characterization of EGF receptor in a water-soluble membrane model system [electronic resource].

Published:: Washington, D.C. : United States. Dept. of Energy. Office of Science, 2017.
Oak Ridge, Tenn. : Distributed by the Office of Scientific and Technical Information, U.S. Dept. of Energy
Physical Description:: Article numbers e0,177,761 : digital, PDF file
Additional Creators:: Lawrence Berkeley National Laboratory, United States. Department of Energy. Office of Science, National Institutes of Health (U.S.), and United States. Department of Energy. Office of Scientific and Technical Information

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Summary:

ErbB (Erythroblastic Leukemia Viral Oncogene Homolog) receptor tyrosine kinases are critical for tissue development and maintenance, and frequently become oncogenic when mutated or overexpressed. In vitro analysis of ErbB receptor kinases can be difficult because of their large size and poor water solubility. Here we report improved production and assembly of the correctly folded full-length EGF receptor (EGFR) into nanolipoprotein particles (NLPs). NLPs are ~10 nm in diameter discoidal cell membrane mimics composed of apolipoproteins surrounding a lipid bilayer. NLPs containing EGFR were synthesized via incubation of baculovirus-produced recombinant EGFR with apolipoprotein and phosphoplipids under conditions that favor self-assembly. The resulting EGFR-NLPs were the correct size, formed dimers and multimers, had intrinsic autophosphorylation activity, and retained the ability to interact with EGFR-targeted ligands and inhibitors consistent with previously-published in vitro binding affinities. Lastly, we anticipate rapid adoption of EGFR-NLPs for structural studies of full-length receptors and drug screening, as well as for the in vitro characterization of ErbB heterodimers and disease-relevant mutants.

Report Numbers:

E 1.99:1395310

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Published through SciTech Connect.
06/06/2017.
PLoS ONE 12 6 ISSN 1932-6203 AM
Tiffany M. Scharadin; Wei He; Yianni Yiannakou; Alexey A. Tomilov; Matthew Saldana; Gino A. Cortopassi; Kermit L. Carraway; Matthew A. Coleman; Paul T. Henderson.

Funding Information:

AC52-07NA27344

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