Relationship between propeptide pH unfolding and inhibitory ability during ProDer p 1 activation mechanism

JOURNAL OF MOLECULAR BIOLOGY

卷:374

Article

Chevigne, Andy ; Barumandzadeh, Roya ; Groslambert, Sylvie ; Cloes, Benoit ; Dehareng, Dominique ; Filee, Patrice ; Marx, Jean-Claude ; Frere, Jean-Marie ; Matagne, Andre ; Jacquet, Alain ; Galleni, Moreno

关键词: allergy;    cysteine protease;    inhibition mechanism;    propeptide;    pH unfolding;   

DOI  :  10.1016/j.jmb.2007.08.025

来源: Elsevier

【 摘 要 】

The major allergen Der p1 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 I 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 propepticles 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 propepticle 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. (c) 2007 Elsevier Ltd. All rights reserved.

【 授权许可】

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