The Crystal Structure of Recombinant proDer p 1, a Major House Dust Mite Proteolytic Allergen

Meno, K.; Thorsted, Peter; Ipsen, Henrik; Kristensen, Ole; Larsen, Jørgen Nedergaard; Spangfort, Michael D.; Gajhede, Michael; Lund, Kaare

doi:10.4049/jimmunol.175.6.3835

Cited by 70 publications

(66 citation statements)

References 50 publications

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“…The three-dimensional coordinates of the initial structure of the propeptide were taken from the crystal structure of the complex between Der p 1 and its propeptide 19 and soaked in a 5-Å layer of water molecules. The geometry was first optimized with a convergence threshold of 0.02 kcal mol − 1 Å − 1 and then submitted to two-dynamics simulations at 300K during 100 ps and 1.1 ns.…”

Section: Molecular Modelingmentioning

confidence: 99%

“…20 Meno et al proposed that the Der p 1 propeptide belongs to a third subfamily characterized by a shorter propeptide with four α-helices and the absence of β-strands. 19 Therefore, the study of the ProDer p 1 activation mechanism could represent an interesting way to highlight the importance of the presence or absence of a conserved propeptide motif and secondary structure elements during the activation of the papain-like precursor.…”

Section: Introductionmentioning

confidence: 99%

“…13,14 Finally, several experiments performed with ProDer p 1 mutants lacking the N-glycosylation sites or with endoglycosylase H-pretreated ProDer p 1 indicated that the glycosylation of recombinant zymogen expressed in P. pastoris can impair the maturation of the allergen. 17,18 Analysis of the X-ray structure of the zymogen form of the allergen reveals that the propeptide of Der p 1 adopts a new fold within the CA1 protease family 19 ( Fig. 1).…”

Section: Introductionmentioning

confidence: 99%

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

Chevigné

Barumandzadeh

Groslambert

et al. 2007

Journal of Molecular Biology

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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.

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Section: Molecular Modelingmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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

Chevigné

Barumandzadeh

Groslambert

et al. 2007

Journal of Molecular Biology

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“…At least 12 isoforms with different amino acid sequences for Der p 1 (18) and two for Der f 1 (12,19) have been identified. Very recently, the crystal structure of a recombinant mutant protein of the Der p 1 precursor was determined (20).…”

mentioning

confidence: 99%

Crucial Commitment of Proteolytic Activity of a Purified Recombinant Major House Dust Mite Allergen Der p1 to Sensitization toward IgE and IgG Responses

Kikuchi

Takai²,

Kuhara³

et al. 2006

The Journal of Immunology

102

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The major proteolytic allergen derived from the house dust mite Dermatophagoides pteronyssinus, Der p1, is one of the most clinically relevant allergens worldwide. In the present study, we evaluate the contribution of the proteolytic activity and structure of a highly purified rDer p 1 to immune responses. Mice were i.p. immunized with three forms of rDer p 1 adsorbed to Alum: one enzymatically active, one treated with an irreversible cysteine protease-specific inhibitor, E-64, and one heat denatured. Immunization with E-64-treated or heat-denatured rDer p 1 elicited much less production of serum total IgE and not only rDer p 1-specific IgE but also IgGs compared with immunization with active rDer p 1. Assays for Ab-binding and its inhibition and structural analyses indicated that E-64-treated rDer p 1 retained its global structure and conformational B cell epitopes. A proliferative response and production of IL-5 by spleen cells restimulated with rDer p 1 were observed on immunization with the active rDer p 1 but not E-64-treated rDer p 1. The cells from mice immunized with heat-denatured rDer p 1 exhibited the highest levels of proliferation and production of IL-5 and IFN-γ. The results indicate that the proteolytic activity of the highly purified rDer p 1 crucially commits to the sensitization process, including both IgE and IgG responses. Additionally, we demonstrated immunogenic differences by functional or structural manipulations of the rDer p 1. The findings have implications for sensitization to this relevant allergen in humans and for the design of modified allergen-vaccines for future allergen-specific immunotherapy.

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“…IgE epitopes of Der f 1 and Der p 1 are dependent on the integrity of the conformational structure (8, 16 -19), and high cross-reactivity between Der f 1 and Der p 1 has been reported (9,11,19,20). The crystal structures of the Der p 1 precursor (21) and mature Der p 1 (22) were determined.…”

mentioning

confidence: 99%

Modulation of Allergenicity of Major House Dust Mite Allergens Der f 1 and Der p 1 by Interaction with an Endogenous Ligand

Takai¹,

Kato²,

Hatanaka

et al. 2009

The Journal of Immunology

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Although many allergens bind endogenous molecules other than Abs in the human body, whether the interaction can modulate allergenicity has been unknown. Here, we investigated the effect of the interaction of recombinant major mite group 1 allergens (Der f 1 and Der p 1), which belong to the papain-like cysteine protease family, with an endogenous protease inhibitor, cystatin A, on their allergenicity. Cystatin A bound reduced forms of the allergens, in which the cysteine residue at the catalytic center of the protease activity was reduced by treatment with l-cysteine, but did not bind oxidized forms. Cystatin A partially inhibited the binding of IgE in mite-allergic volunteers’ sera to the reduced forms, but unexpectedly enhanced the basophil histamine-releasing activity. A catalytic site-mutant of Der f 1 behaved in terms of histamine release, similarly to the reduced form. Molecular modeling showed that cystatin A interacts with the allergens within a narrow area. The results indicate that interaction with cystatin A reduces the limited number of IgE epitopes of the allergens but enhances their biological activity to release histamine, suggesting a new concept, that interaction between allergens and their endogenous ligands modulates the allergenicity even toward enhancement in the effector phase. On the other hand, i.p. immunization without alum of mice with cystatin A-treated reduced Der f 1 induced less serum Der f 1-specific IgE than immunization with reduced Der f 1 alone, suggesting that endogenous protease inhibitors suppress the induction of allergen-specific IgE, which is dependent on the enzymatic activity of cysteine protease-allergens, in the sensitization process.

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The Crystal Structure of Recombinant proDer p 1, a Major House Dust Mite Proteolytic Allergen

Cited by 70 publications

References 50 publications

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

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

Crucial Commitment of Proteolytic Activity of a Purified Recombinant Major House Dust Mite Allergen Der p1 to Sensitization toward IgE and IgG Responses

Modulation of Allergenicity of Major House Dust Mite Allergens Der f 1 and Der p 1 by Interaction with an Endogenous Ligand

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