Synthesis of PAF, an antifungal protein from P. chrysogenum, by native chemical ligation: Native disulfide pattern and fold obtained upon oxidative refolding

Györgyi Váradi, Gábor K. Tõth, Zoltán Kele, Lászlõ Galgõczy, Ádám Fizil, Gyula Batta

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

The folding of disulfide proteins is of considerable interest because knowledge of this may influence our present understanding of protein folding. However, sometimes even the disulfide pattern cannot be unequivocally determined by the available experimental techniques. For example, the structures of a few small antifungal proteins (PAF, AFP) have been disclosed recently using NMR spectroscopy but with some ambiguity in the actual disulfide pattern. For this reason, we carried out the chemical synthesis of PAF. Probing different approaches, the oxidative folding of the synthetic linear PAF yielded a folded protein that has identical structure and antifungal activity as the native PAF. In contrast, unfolded linear PAF was inactive, a result that may have implications concerning its redox state in the mode of action. Thiol soul mates: PAF, an antifungal protein from P.a chrysogenum, can be prepared synthetically by using a combination of solid-phase peptide synthesis (SPPS) and native chemical ligation followed by oxidative folding (see scheme; Boc=tert-butoxycarbonyl, Fmoc=9-fluorenylmethoxycarbonyl, MBHA= methylbenzhydrylamine). Using this method, PAF is prepared with the correct disulfide-bond connectivity and fold.

Original languageEnglish
Pages (from-to)12684-12692
Number of pages9
JournalChemistry - A European Journal
Volume19
Issue number38
DOIs
Publication statusPublished - Sep 16 2013

Keywords

  • NMR spectroscopy
  • antifungal agents
  • peptides
  • protein folding
  • solid-phase synthesis

ASJC Scopus subject areas

  • Catalysis
  • Organic Chemistry

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