Cloning an artificial gene encoding angiostatic anginex: From designed peptide to functional recombinant protein

Ricardo J.M.G.E. Brandwijk, Irina Nesmelova, Ruud P.M. Dings, Kevin H. Mayo, Victor L.J.L. Thijssen, Arjan W. Griffioen

Research output: Contribution to journalArticlepeer-review

20 Scopus citations

Abstract

Anginex, a designed peptide 33-mer, is a potent angiogenesis inhibitor and anti-tumor agent in vivo. Anginex functions by inhibiting endothelial cell (EC) proliferation and migration leading to detachment and apoptosis of activated EC's. To better understand tumor endothelium targeting properties of anginex and enable its use in gene therapy, we constructed an artificial gene encoding the biologically exogenous peptide and produced the protein recombinantly in Pichia pastoris. Mass spectrometry shows recombinant anginex to be a dimer and circular dichroism shows the recombinant protein folds with β-strand structure like the synthetic peptide. Moreover, like parent anginex, the recombinant protein is active at inhibiting EC growth and migration, as well as inhibiting angiogenesis in vivo in the chorioallantoic membrane of the chick embryo. This study demonstrated that it is possible to produce a functionally active protein version of a rationally designed peptide, using an artificial gene and the recombinant protein approach.

Original languageEnglish (US)
Pages (from-to)1261-1268
Number of pages8
JournalBiochemical and Biophysical Research Communications
Volume333
Issue number4
DOIs
StatePublished - Aug 12 2005

Bibliographical note

Funding Information:
This study was supported by generous research grants from the Technology Foundation STW, applied science division of NWO and the technology program of the Ministry of Economic Affairs (MPG 5465 to A.W.G.), and from the National Institutes of Health (CA-96090 to K.H.M.). We are grateful to Denisha Walek for expert technical assistance.

Keywords

  • Anginex
  • Angiogenesis
  • Endothelial cells
  • Functional characterization
  • Yeast expression system

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