The Lys-Asp-Tyr triad within the mite allergen der p 1 propeptide is a critical structural element for the pH-dependent initiation of the protease maturation

Andy Chevigné, Vincenzo Campizi, Martyna Szpakowska, David Bourry, Marie Eve Dumez, José C. Martins, André Matagne, Moreno Galleni, Alain Jacquet*

*Corresponding author for this work

Research output: Contribution to journalArticleResearchpeer-review

4 Citations (Scopus)

Abstract

The major house dust mite allergen, Der p 1, is a papain-like cysteine protease expressed as an inactive precursor, proDer p 1, carrying an N-terminal propeptide with a unique structure. The maturation of the zymogen into an enzymatically-active form of Der p 1 is a multistep autocatalytic process initiated under acidic conditions through conformational changes of the propeptide, leading to the loss of its inhibitory ability and its subsequent gradual cleavage. The aims of this study were to characterize the residues present in the Der p 1 propeptide involved in the initiation of the zymogen maturation process, but also to assess the impact of acidic pH on the propeptide structure, the activity of Der p 1 and the fate of the propeptide. Using various complementary enzymatic and structural approaches, we demonstrated that a structural triad K17p-D51p-Y19p within the N-terminal domain of the propeptide is essential for its stabilization and the sensing of pH changes. Particularly, the protonation of D51p under acidic conditions unfolds the propeptide through disruption of the K17p-D51p salt bridge, reduces its inhibition capacity and unmasks the buried residues K17p and Y19p constituting the first maturation cleavage site of the zymogen. Our results also evidenced that this triad acts in a cooperative manner with other propeptide pH-responsive elements, including residues E56p and E80p, to promote the propeptide unfolding and/or to facilitate its proteolysis. Furthermore, we showed that acidic conditions modify Der p 1 proteolytic specificity and confirmed that the formation of the first intermediate represents the limiting step of the in vitro Der p 1 maturation process. Altogether, our results provide new insights into the early events of the mechanism of proDer p 1 maturation and identify a unique structural triad acting as a stabilizing and a pH-sensing regulatory element.

Original languageEnglish
Article number1087
JournalInternational Journal of Molecular Sciences
Volume18
Issue number5
DOIs
Publication statusPublished - 20 May 2017

Keywords

  • Cysteine protease
  • Der p 1
  • Maturation
  • Propeptide
  • pH sensor
  • pH unfolding

Fingerprint

Dive into the research topics of 'The Lys-Asp-Tyr triad within the mite allergen der p 1 propeptide is a critical structural element for the pH-dependent initiation of the protease maturation'. Together they form a unique fingerprint.

Cite this