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The proline-rich motif of the proDer p 3 allergen propeptide is crucial for protease-protease interaction

(2013) PLOS ONE. 8(9).
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Abstract
The majority of proteases are synthesized in an inactive form, termed zymogen, which consists of a propeptide and a protease domain. The propeptide is commonly involved in the correct folding and specific inhibition of the enzyme. The propeptide of the house dust mite allergen Der p 3, NPILPASPNAT, contains a proline-rich motif (PRM), which is unusual for a trypsin-like protease. By truncating the propeptide or replacing one or all of the prolines in the non-glycosylated zymogen with alanine(s), we demonstrated that the full-length propeptide is not required for correct folding and thermal stability and that the PRM is important for the resistance of proDer p 3 to undesired proteolysis when the protein is expressed in Pichia pastoris. Additionally, we followed the maturation time course of proDer p 3 by coupling a quenched-flow assay to mass spectrometry analysis. This approach allowed to monitor the evolution of the different species and to determine the steady-state kinetic parameters for activation of the zymogen by the major allergen Der p 1. This experiment demonstrated that prolines 5 and 8 are crucial for proDer p 3-Der p 1 interaction and for activation of the zymogen.
Keywords
ZYMOGEN ACTIVATION, INTRAMOLECULAR CHAPERONE, DUST-MITE ALLERGEN, GLYCOSYLATION, PEPTIDES, SUBTILISIN, RECOMBINANT, IGE, ACTIVE-SITE, TRYPSIN

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Chicago
Dumez, Marie-Eve, Julie Herman, Vincenzo Campisi, Ahlem Bouaziz, Frédéric Rosu, André Luxen, Isabel Vandenberghe, et al. 2013. “The Proline-rich Motif of the proDer p 3 Allergen Propeptide Is Crucial for Protease-protease Interaction.” Plos One 8 (9).
APA
Dumez, M.-E., Herman, J., Campisi, V., Bouaziz, A., Rosu, F., Luxen, A., Vandenberghe, I., et al. (2013). The proline-rich motif of the proDer p 3 allergen propeptide is crucial for protease-protease interaction. PLOS ONE, 8(9).
Vancouver
1.
Dumez M-E, Herman J, Campisi V, Bouaziz A, Rosu F, Luxen A, et al. The proline-rich motif of the proDer p 3 allergen propeptide is crucial for protease-protease interaction. PLOS ONE. 2013;8(9).
MLA
Dumez, Marie-Eve, Julie Herman, Vincenzo Campisi, et al. “The Proline-rich Motif of the proDer p 3 Allergen Propeptide Is Crucial for Protease-protease Interaction.” PLOS ONE 8.9 (2013): n. pag. Print.
@article{5659177,
  abstract     = {The majority of proteases are synthesized in an inactive form, termed zymogen, which consists of a propeptide and a protease domain. The propeptide is commonly involved in the correct folding and specific inhibition of the enzyme. The propeptide of the house dust mite allergen Der p 3, NPILPASPNAT, contains a proline-rich motif (PRM), which is unusual for a trypsin-like protease. By truncating the propeptide or replacing one or all of the prolines in the non-glycosylated zymogen with alanine(s), we demonstrated that the full-length propeptide is not required for correct folding and thermal stability and that the PRM is important for the resistance of proDer p 3 to undesired proteolysis when the protein is expressed in Pichia pastoris. Additionally, we followed the maturation time course of proDer p 3 by coupling a quenched-flow assay to mass spectrometry analysis. This approach allowed to monitor the evolution of the different species and to determine the steady-state kinetic parameters for activation of the zymogen by the major allergen Der p 1. This experiment demonstrated that prolines 5 and 8 are crucial for proDer p 3-Der p 1 interaction and for activation of the zymogen.},
  articleno    = {e68014},
  author       = {Dumez, Marie-Eve and Herman, Julie and Campisi, Vincenzo and Bouaziz, Ahlem and Rosu, Fr{\'e}d{\'e}ric and Luxen, Andr{\'e} and Vandenberghe, Isabel and de Pauw, Edwin and Fr{\`e}re, Jean-Marie and Matagne, Andr{\'e} and Chevign{\'e}, Andy and Galleni, Moreno},
  issn         = {1932-6203},
  journal      = {PLOS ONE},
  keyword      = {ZYMOGEN ACTIVATION,INTRAMOLECULAR CHAPERONE,DUST-MITE ALLERGEN,GLYCOSYLATION,PEPTIDES,SUBTILISIN,RECOMBINANT,IGE,ACTIVE-SITE,TRYPSIN},
  language     = {eng},
  number       = {9},
  pages        = {12},
  title        = {The proline-rich motif of the proDer p 3 allergen propeptide is crucial for protease-protease interaction},
  url          = {http://dx.doi.org/10.1371/journal.pone.0068014},
  volume       = {8},
  year         = {2013},
}

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