Toward the semisynthesis of multidomain transmembrane receptors: Modification of Eph tyrosine kinases

Nikhil Singla, Juha Pekka Himanen, Tom W. Muir, Dimitar B. Nikolov

Research output: Contribution to journalArticlepeer-review

11 Scopus citations

Abstract

Expressed protein ligation (EPL) is a protein engineering approach that allows the modification or assembly of a target protein from multiple recombinant and synthetic polypeptides. EPL has been previously used to modify intracellular proteins and small integral membrane proteins for structural and functional studies. Here we describe the semisynthetic site-specific modification of the complete, multidomain extracellular regions of both A and B classes of Eph receptor tyrosine kinases. We show that the ectodomains of these receptors can be ligated to different peptides under carefully established experimental conditions, while their biological activity is retained. This work extends the boundaries of the EPL technique for semisynthesis of multidomain, extracellular, disulfide-bonded, and glycosylated proteins and highlights its potential application for reconstituting entire single-pass transmembrane proteins.

Original languageEnglish (US)
Pages (from-to)1740-1747
Number of pages8
JournalProtein Science
Volume17
Issue number10
DOIs
StatePublished - Oct 2008
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology

Keywords

  • Eph receptor tyrosine kinase
  • Ephrins
  • Expressed protein ligation
  • Inteins
  • Multidomain extracellular region
  • Peptides

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