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Title: Cell-free expression of functional receptor tyrosine kinases

Abstract

Receptor tyrosine kinases (RTKs) play critical roles in physiological and pathological processes, and are important anticancer drug targets. In vitro mechanistic and drug discovery studies of full-length RTKs require protein that is both fully functional and free from contaminating proteins. Here we describe a rapid cell-free and detergent-free co-translation method for producing full-length and functional ERBB2 and EGFR receptor tyrosine kinases supported by water-soluble apolipoprotein A-I based nanolipoprotein particles.

Authors:
 [1];  [1];  [1];  [1];  [1];  [1];  [2];  [3];  [4];  [4];  [5]
  1. Univ. of California Davis School of Medicine, Davis, CA (United States)
  2. Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Univ. of California, Santa Cruz, CA (United States)
  3. Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Scripps Research Inst., La Jolla, CA (United States)
  4. Univ. of California Davis School of Medicine, Davis, CA (United States); Univ. of California Davis Comprehensive Cancer Center, Sacramento, CA (United States)
  5. Univ. of California Davis School of Medicine, Davis, CA (United States); Univ. of California Davis Comprehensive Cancer Center, Sacramento, CA (United States); Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States)
Publication Date:
Research Org.:
Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States)
Sponsoring Org.:
USDOE National Nuclear Security Administration (NNSA)
OSTI Identifier:
1515361
Report Number(s):
LLNL-JRNL-741146
Journal ID: ISSN 2045-2322; 895483
Grant/Contract Number:  
AC52-07NA27344
Resource Type:
Accepted Manuscript
Journal Name:
Scientific Reports
Additional Journal Information:
Journal Volume: 5; Journal Issue: 1; Journal ID: ISSN 2045-2322
Publisher:
Nature Publishing Group
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES

Citation Formats

He, Wei, Scharadin, Tiffany M., Saldana, Matthew, Gellner, Candice, Hoang-Phou, Steven, Takanishi, Christina, Hura, Gregory L., Tainer, John A., Carraway III, Kermit L., Henderson, Paul T., and Coleman, Matthew A. Cell-free expression of functional receptor tyrosine kinases. United States: N. p., 2015. Web. doi:10.1038/srep12896.
He, Wei, Scharadin, Tiffany M., Saldana, Matthew, Gellner, Candice, Hoang-Phou, Steven, Takanishi, Christina, Hura, Gregory L., Tainer, John A., Carraway III, Kermit L., Henderson, Paul T., & Coleman, Matthew A. Cell-free expression of functional receptor tyrosine kinases. United States. doi:10.1038/srep12896.
He, Wei, Scharadin, Tiffany M., Saldana, Matthew, Gellner, Candice, Hoang-Phou, Steven, Takanishi, Christina, Hura, Gregory L., Tainer, John A., Carraway III, Kermit L., Henderson, Paul T., and Coleman, Matthew A. Fri . "Cell-free expression of functional receptor tyrosine kinases". United States. doi:10.1038/srep12896. https://www.osti.gov/servlets/purl/1515361.
@article{osti_1515361,
title = {Cell-free expression of functional receptor tyrosine kinases},
author = {He, Wei and Scharadin, Tiffany M. and Saldana, Matthew and Gellner, Candice and Hoang-Phou, Steven and Takanishi, Christina and Hura, Gregory L. and Tainer, John A. and Carraway III, Kermit L. and Henderson, Paul T. and Coleman, Matthew A.},
abstractNote = {Receptor tyrosine kinases (RTKs) play critical roles in physiological and pathological processes, and are important anticancer drug targets. In vitro mechanistic and drug discovery studies of full-length RTKs require protein that is both fully functional and free from contaminating proteins. Here we describe a rapid cell-free and detergent-free co-translation method for producing full-length and functional ERBB2 and EGFR receptor tyrosine kinases supported by water-soluble apolipoprotein A-I based nanolipoprotein particles.},
doi = {10.1038/srep12896},
journal = {Scientific Reports},
number = 1,
volume = 5,
place = {United States},
year = {2015},
month = {8}
}

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Cited by: 13 works
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Works referenced in this record:

Applications of small-angle X-ray scattering to biomacromolecular solutions
journal, February 2013

  • Petoukhov, Maxim V.; Svergun, Dmitri I.
  • The International Journal of Biochemistry & Cell Biology, Vol. 45, Issue 2
  • DOI: 10.1016/j.biocel.2012.10.017

Characterizing diffusion dynamics of a membrane protein associated with nanolipoproteins using fluorescence correlation spectroscopy
journal, January 2011

  • Gao, Tingjuan; Blanchette, Craig D.; He, Wei
  • Protein Science, Vol. 20, Issue 2
  • DOI: 10.1002/pro.577

Cell-free Co-expression of Functional Membrane Proteins and Apolipoprotein, Forming Soluble Nanolipoprotein Particles
journal, July 2008

  • Cappuccio, Jenny A.; Blanchette, Craig D.; Sulchek, Todd A.
  • Molecular & Cellular Proteomics, Vol. 7, Issue 11
  • DOI: 10.1074/mcp.M800191-MCP200

Small-angle scattering gives direct structural information about a membrane protein inside a lipid environment
journal, January 2014

  • Kynde, Søren A. R.; Skar-Gislinge, Nicholas; Pedersen, Martin Cramer
  • Acta Crystallographica Section D Biological Crystallography, Vol. 70, Issue 2
  • DOI: 10.1107/S1399004713028344

Conformational Coupling across the Plasma Membrane in Activation of the EGF Receptor
journal, January 2013


Novel anticancer targets: revisiting ERBB2 and discovering ERBB3
journal, June 2009

  • Baselga, José; Swain, Sandra M.
  • Nature Reviews Cancer, Vol. 9, Issue 7
  • DOI: 10.1038/nrc2656

Finding the missing links in EGFR
journal, January 2012

  • Bessman, Nicholas J.; Lemmon, Mark A.
  • Nature Structural & Molecular Biology, Vol. 19, Issue 1
  • DOI: 10.1038/nsmb.2221

Cell-free protein synthesis: Applications come of age
journal, September 2012


Fluorescent-labeled growth factor molecules serve as probes for receptor binding and endocytosis
journal, November 1993

  • Carraway, Kermit L.; Cerione, Richard A.
  • Biochemistry, Vol. 32, Issue 45
  • DOI: 10.1021/bi00096a014

The Cell-Free Integration of a Polytopic Mitochondrial Membrane Protein into Liposomes Occurs Cotranslationally and in a Lipid-Dependent Manner
journal, September 2012


Intragenic ERBB2 kinase mutations in tumours
journal, September 2004

  • Stephens, Philip; Hunter, Chris; Bignell, Graham
  • Nature, Vol. 431, Issue 7008
  • DOI: 10.1038/431525b

Comprehensive macromolecular conformations mapped by quantitative SAXS analyses
journal, April 2013

  • Hura, Greg L.; Budworth, Helen; Dyer, Kevin N.
  • Nature Methods, Vol. 10, Issue 6
  • DOI: 10.1038/nmeth.2453

Robust, high-throughput solution structural analyses by small angle X-ray scattering (SAXS)
journal, July 2009

  • Hura, Greg L.; Menon, Angeli L.; Hammel, Michal
  • Nature Methods, Vol. 6, Issue 8
  • DOI: 10.1038/nmeth.1353

A monomeric G protein-coupled receptor isolated in a high-density lipoprotein particle efficiently activates its G protein
journal, April 2007

  • Whorton, M. R.; Bokoch, M. P.; Rasmussen, S. G. F.
  • Proceedings of the National Academy of Sciences, Vol. 104, Issue 18, p. 7682-7687
  • DOI: 10.1073/pnas.0611448104

Characterization of De Novo Synthesized GPCRs Supported in Nanolipoprotein Discs
journal, September 2012


Review of epidermal growth factor receptor biology
journal, June 2004


Human breast cancer: correlation of relapse and survival with amplification of the HER-2/neu oncogene
journal, January 1987


Untangling the ErbB signalling network
journal, February 2001

  • Yarden, Yosef; Sliwkowski, Mark X.
  • Nature Reviews Molecular Cell Biology, Vol. 2, Issue 2
  • DOI: 10.1038/35052073

An Allosteric Mechanism for Activation of the Kinase Domain of Epidermal Growth Factor Receptor
journal, June 2006


Functional and Structural Stability of the Epidermal Growth Factor Receptor in Detergent Micelles and Phospholipid Nanodiscs
journal, September 2008

  • Mi, Li-Zhi; Grey, Michael J.; Nishida, Noritaka
  • Biochemistry, Vol. 47, Issue 39
  • DOI: 10.1021/bi801006s

Epidermal growth factor stimulates the phosphorylation of synthetic tyrosine-containing peptides by A431 cell membranes.
journal, March 1982

  • Pike, L. J.; Gallis, B.; Casnellie, J. E.
  • Proceedings of the National Academy of Sciences, Vol. 79, Issue 5
  • DOI: 10.1073/pnas.79.5.1443

Self-assembly of single integral membrane proteins into soluble nanoscale phospholipid bilayers
journal, November 2003

  • Bayburt, Timothy H.; Sligar, Stephen G.
  • Protein Science, Vol. 12, Issue 11, p. 2476-2481
  • DOI: 10.1110/ps.03267503

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