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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 UGent, Marie-Eve Dumez, José Martins UGent, André Matagne, Moreno Galleni and Alain Jacquet (2017) INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES. 18(5).
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.
Please use this url to cite or link to this publication:
author
organization
year
type
journalArticle (original)
publication status
published
subject
keyword
HOUSE-DUST-MITE, PROCATHEPSIN-L, DERMATOPHAGOIDES-PTERONYSSINUS, RECOMBINANT PRO-DER-P-1, ACTIVATION MECHANISM, PROTEOLYTIC ACTIVITY, CYSTEINE PROTEASES, PICHIA-PASTORIS, INNATE IMMUNITY, CATHEPSIN K, cysteine protease, Der p 1, pH sensor, pH unfolding, propeptide, maturation
journal title
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
Int. J. Mol. Sci.
volume
18
issue
5
article number
1087
pages
16 pages
Web of Science type
Article
Web of Science id
000404113900190
ISSN
1422-0067
DOI
10.3390/ijms18051087
language
English
UGent publication?
yes
classification
A1
copyright statement
Creative Commons Attribution 4.0 International Public License (CC-BY 4.0)
id
8534803
handle
http://hdl.handle.net/1854/LU-8534803
date created
2017-10-19 13:12:21
date last changed
2017-11-17 14:58:29
@article{8534803,
  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.},
  articleno    = {1087},
  author       = {Chevign{\'e}, Andy and Campizi, Vincenzo and Szpakowska, Martyna and Bourry, David and Dumez, Marie-Eve and Martins, Jos{\'e} and Matagne, Andr{\'e} and Galleni, Moreno and Jacquet, Alain},
  issn         = {1422-0067},
  journal      = {INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES},
  keyword      = {HOUSE-DUST-MITE,PROCATHEPSIN-L,DERMATOPHAGOIDES-PTERONYSSINUS,RECOMBINANT PRO-DER-P-1,ACTIVATION MECHANISM,PROTEOLYTIC ACTIVITY,CYSTEINE PROTEASES,PICHIA-PASTORIS,INNATE IMMUNITY,CATHEPSIN K,cysteine protease,Der p 1,pH sensor,pH unfolding,propeptide,maturation},
  language     = {eng},
  number       = {5},
  pages        = {16},
  title        = {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},
  url          = {http://dx.doi.org/10.3390/ijms18051087},
  volume       = {18},
  year         = {2017},
}

Chicago
Chevigné, Andy, Vincenzo Campizi, Martyna Szpakowska, David Bourry, Marie-Eve Dumez, José Martins, André Matagne, Moreno Galleni, and Alain Jacquet. 2017. “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.” International Journal of Molecular Sciences 18 (5).
APA
Chevigné, A., Campizi, V., Szpakowska, M., Bourry, D., Dumez, M.-E., Martins, J., Matagne, A., et al. (2017). 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. INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES, 18(5).
Vancouver
1.
Chevigné A, Campizi V, Szpakowska M, Bourry D, Dumez M-E, Martins J, et al. 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. INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES. 2017;18(5).
MLA
Chevigné, Andy, Vincenzo Campizi, Martyna Szpakowska, et al. “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.” INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES 18.5 (2017): n. pag. Print.