Relationship between Propeptide pH Unfolding and Inhibitory Ability during ProDer p 1 Activation Mechanism

Andy Chevigné, Roya Barumandzadeh, Sylvie Groslambert, Benoît Cloes, Dominique Dehareng, Patrice Filée, Jean Claude Marx, Jean Marie Frère, André Matagne, Alain Jacquet, Moreno Galleni*

*Corresponding author for this work

Research output: Contribution to journalArticleResearchpeer-review

23 Citations (Scopus)

Abstract

The major allergen Der p 1 of the house dust mite Dermatophagoides pteronyssinus is a papain-like cysteine protease (CA1) produced as an inactive precursor and associated with allergic diseases. The propeptide of Der p 1 exhibits a specific fold that makes it unique in the CA1 propeptide family. In this study, we investigated the activation steps involved in the maturation of the recombinant protease Der p 1 expressed in Pichia pastoris and the interaction of the full-length and truncated soluble propeptides with their parent enzyme in terms of activity inhibition and BIAcore interaction analysis. According to our results, the activation of protease Der p 1 is a multistep mechanism that is characterized by at least two intermediates. The propeptide strongly inhibits unglycosylated and glycosylated recombinant Der p 1 (KD = 7 nM) at neutral pH. This inhibition is pH dependent. It decreases from pH 7 to pH 4 and can be related to conformational changes of the propeptide characterized by an increase of its flexibility and formation of a molten globule state. Our results indicate that activation of the zymogen at pH 4 is a compromise between activity preservation and propeptide unfolding.

Original languageEnglish
Pages (from-to)170-185
Number of pages16
JournalJournal of Molecular Biology
Volume374
Issue number1
DOIs
Publication statusPublished - 16 Nov 2007
Externally publishedYes

Keywords

  • allergy
  • cysteine protease
  • inhibition mechanism
  • pH unfolding
  • propeptide

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