Synthesis and biochemical characterization of EGF receptor in a water-soluble membrane model system

Tiffany M. Scharadin, Wei He, Yianni Yiannakou, Alexey Tomilov, Matthew Saldana, Gino A Cortopassi, Kermit L Carraway, Matthew A Coleman, Paul Henderson

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

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. 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.

Original languageEnglish (US)
Article number0177761
JournalPLoS One
Volume12
Issue number6
DOIs
StatePublished - Jun 1 2017

Fingerprint

Epidermal Growth Factor Receptor
Membranes
synthesis
Water
Apolipoproteins
apolipoproteins
water
receptors
phosphotransferases (kinases)
Preclinical Drug Evaluations
Baculoviridae
Drug Receptors
Receptor Protein-Tyrosine Kinases
Lipid Bilayers
lipid bilayers
Lipid bilayers
Oncogenes
oncogenes
protein phosphorylation
Solubility

ASJC Scopus subject areas

  • Medicine(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

Cite this

Synthesis and biochemical characterization of EGF receptor in a water-soluble membrane model system. / Scharadin, Tiffany M.; He, Wei; Yiannakou, Yianni; Tomilov, Alexey; Saldana, Matthew; Cortopassi, Gino A; Carraway, Kermit L; Coleman, Matthew A; Henderson, Paul.

In: PLoS One, Vol. 12, No. 6, 0177761, 01.06.2017.

Research output: Contribution to journalArticle

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AU - Cortopassi, Gino A

AU - Carraway, Kermit L

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