Reference : The proline-rich motif of the proDer p 3 allergen propeptide is crucial for protease-...
Scientific journals : Article
Life sciences : Biochemistry, biophysics & molecular biology
http://hdl.handle.net/2268/160740
The proline-rich motif of the proDer p 3 allergen propeptide is crucial for protease-protease interaction.
English
Dumez, Marie-Eve mailto [Université de Liège - ULiège > Département des sciences de la vie > Macromolécules biologiques >]
Herman, Julie [Université de Liège - ULiège > Sciences de la Vie > Macromolécules Biologiques, C.I.P. > >]
Campisi, Vincenzo [Université de Liège - ULiège > Département des sciences de la vie > Macromolécules biologiques >]
Bouaziz, Ahlem mailto [Université de Liège - ULiège > Sciences de la Vie > Macromolécules Biologiques, C.I.P. > >]
Rosu, Frederic [> >]
Luxen, André mailto [Université de Liège - ULiège > Département de chimie (sciences) > Chimie organique de synthèse >]
Vandenberghe, Isabel [> >]
De Pauw, Edwin mailto [Université de Liège - ULiège > Département de chimie (sciences) > GIGA-R : Laboratoire de spectrométrie de masse (L.S.M.) >]
Frère, Jean-Marie mailto [Université de Liège - ULiège > > Centre d'ingénierie des protéines >]
Matagne, André mailto [Université de Liège - ULiège > Département des sciences de la vie > Enzymologie et repliement des protéines >]
Chevigne, Andy [> >]
Galleni, Moreno mailto [Université de Liège - ULiège > Département des sciences de la vie > Macromolécules biologiques >]
2013
PLoS ONE
8
9
e68014
Yes (verified by ORBi)
International
1932-6203
United States
[en] serine protease ; trypsin ; fluorescence ; thermal unfolding ; enzyme kinetics ; mass spectrometry
[en] 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.
http://hdl.handle.net/2268/160740
10.1371/journal.pone.0068014

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