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

doi:10.1016/j.jmb.2007.08.025

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 journal › Article › Research › peer-review

28 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 (K_D = 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 language	English
Pages (from-to)	170-185
Number of pages	16
Journal	Journal of Molecular Biology
Volume	374
Issue number	1
DOIs	https://doi.org/10.1016/j.jmb.2007.08.025
Publication status	Published - 16 Nov 2007
Externally published	Yes

Keywords

allergy
cysteine protease
inhibition mechanism
pH unfolding
propeptide

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10.1016/j.jmb.2007.08.025

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@article{284e038dfaf74a028289189c58008fec,

title = "Relationship between Propeptide pH Unfolding and Inhibitory Ability during ProDer p 1 Activation Mechanism",

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.",

keywords = "allergy, cysteine protease, inhibition mechanism, pH unfolding, propeptide",

author = "Andy Chevign{\'e} and Roya Barumandzadeh and Sylvie Groslambert and Beno{\^i}t Cloes and Dominique Dehareng and Patrice Fil{\'e}e and Marx, {Jean Claude} and Fr{\`e}re, {Jean Marie} and Andr{\'e} Matagne and Alain Jacquet and Moreno Galleni",

note = "Funding Information: This work was supported by the Belgian National Fund for Scientific Research (FNRS), the Walloon Region (Direction G{\'e}n{\'e}rale des Technologies, de la Recherche et de l'Energie) of Belgium (Vaccin 215133 and Allervac 616293) and the Fonds de la Recherche Fondamentale et Collective (2.4.524.03, 2.4.511.06 and 9.4519.98). A.C. is an aspirant of the FNRS (Brussels, Belgium). A.M. is a research associate of the FNRS and is supported in part by a grant from the Fonds de la Recherche Fondamentale et Collective (contract no. 2.4550.05). We thank Marie-Eve Dumez for its helpful comments, Marc Vanhove for its helpful support on BIAcore analysis, Nicole Otthiers for performing the N-terminal sequencing and GlaxoSmithKline Belgium for providing access to ProDer p 1 DNA sequences. ",

year = "2007",

month = nov,

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doi = "10.1016/j.jmb.2007.08.025",

language = "English",

volume = "374",

pages = "170--185",

journal = "Journal of Molecular Biology",

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T1 - Relationship between Propeptide pH Unfolding and Inhibitory Ability during ProDer p 1 Activation Mechanism

AU - Chevigné, Andy

AU - Barumandzadeh, Roya

AU - Groslambert, Sylvie

AU - Cloes, Benoît

AU - Dehareng, Dominique

AU - Filée, Patrice

AU - Marx, Jean Claude

AU - Frère, Jean Marie

AU - Matagne, André

AU - Jacquet, Alain

AU - Galleni, Moreno

N1 - Funding Information: This work was supported by the Belgian National Fund for Scientific Research (FNRS), the Walloon Region (Direction Générale des Technologies, de la Recherche et de l'Energie) of Belgium (Vaccin 215133 and Allervac 616293) and the Fonds de la Recherche Fondamentale et Collective (2.4.524.03, 2.4.511.06 and 9.4519.98). A.C. is an aspirant of the FNRS (Brussels, Belgium). A.M. is a research associate of the FNRS and is supported in part by a grant from the Fonds de la Recherche Fondamentale et Collective (contract no. 2.4550.05). We thank Marie-Eve Dumez for its helpful comments, Marc Vanhove for its helpful support on BIAcore analysis, Nicole Otthiers for performing the N-terminal sequencing and GlaxoSmithKline Belgium for providing access to ProDer p 1 DNA sequences.

PY - 2007/11/16

Y1 - 2007/11/16

N2 - 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.

AB - 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.

KW - allergy

KW - cysteine protease

KW - inhibition mechanism

KW - pH unfolding

KW - propeptide

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DO - 10.1016/j.jmb.2007.08.025

M3 - Article

C2 - 17916363

AN - SCOPUS:35348956380

SN - 0022-2836

VL - 374

SP - 170

EP - 185

JO - Journal of Molecular Biology

JF - Journal of Molecular Biology

IS - 1

ER -

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

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