High-level expression of a full-length Eph receptor Journal Article


Authors: Paavilainen, S.; Grandy, D.; Karelehto, E.; Chang, E.; Susi, P.; Erdjument-Bromage, H.; Nikolov, D.; Himanen, J.
Article Title: High-level expression of a full-length Eph receptor
Abstract: Eph receptors are the largest family of Receptor Tyrosine Kinases containing a single membrane-spanning segment. They are involved in a various developmental and cell-cell communication events. Although there is extensive structural information available on both the extra- and intracellular regions of Eph's in isolation, no structures are available for the entire receptor. To facilitate structural studies on functionally relevant Eph/ephrin complexes, we have developed an expression system for producing the full-length human EphA2 receptor. We successfully expressed milligram amounts of the receptor using baculovirus-based vector and insect cells. We were also able to extract the protein from the cell membranes and purify it to near homogeneity in two simple steps. The purified receptor was shown to retain its biological activity in terms of both binding to its functional ligands and being able to auto-phosphorylate the key tyrosine residues of the cytoplasmic kinase domain. © 2013 Published by Elsevier Inc.
Keywords: tyrosine; receptor; eph receptors; overexpression; kinases; insect cell; transmembrane proteins
Journal Title: Protein Expression and Purification
Volume: 92
Issue: 1
ISSN: 1046-5928
Publisher: Academic Press Inc., Elsevier Science  
Date Published: 2013-11-01
Start Page: 112
End Page: 118
Language: English
DOI: 10.1016/j.pep.2013.08.016
PROVIDER: scopus
PUBMED: 24036371
PMCID: PMC3919533
DOI/URL:
Notes: --- - "Export Date: 1 November 2013" - "CODEN: PEXPE" - "Source: Scopus"
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MSK Authors
  1. Dimitar B Nikolov
    84 Nikolov
  2. Juha P Himanen
    49 Himanen
  3. David U Grandy
    1 Grandy
  4. Elizabeth Chang
    3 Chang