Maturation of the activities of recombinant mite allergens Der p 1 and Der f 1, and its implication in the blockade of proteolytic activity

Takai, Toshiro; Mineki, Reiko; Nakazawa, Takahiro; Takaoka, Masatoshi; Yasueda, Hiroshi; Murayama, Kimie; Okumura, Ko; Ogawa, Hideoki

doi:10.1016/s0014-5793(02)03534-2

Cited by 44 publications

(85 citation statements)

References 49 publications

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“…However, surprisingly, for the N16pQ and N16pQ/N52Q mutants, partial activation occurred during the production, leading to the appearance of different forms. The N-terminal sequencing of the purified mutants revealed the presence of at least three forms, previously described by Takai et al and Jacquet et al 13,14 The first one displayed the ATFE sequence, which corresponds to the cleavage of the peptide bond between Tyr19p and Ala20p. This site is only three residues away from the propeptide Nglycosylation site and gives rise to the loss of the first α-helix of the propeptide (Fig.…”

Section: Resultsmentioning

confidence: 62%

“…[12][13][14] Despite several attempts, in vitro demonstration of the activation of ProDer p 1 remains partly unsuccessful. For example, purified recombinant proforms of Der p 1 expressed in mammalian CHO and insect Drosophila cells could be converted into mature forms by incubation at pH 4 but do not exhibit enzymatic activity.…”

Section: Introductionmentioning

confidence: 99%

“…15,16 More recently, however, several groups have succeeded in converting ProDer p 1 expressed in Pichia pastoris into mature enzymatically active forms with IgE reactivity. 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.…”

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: Resultsmentioning

confidence: 62%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

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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“…Generation of the Der p 1 and Der f 1 Protein-Recombinant Der p 1 and Der f 1 proteins were generated as described before (33,34). Briefly, proforms of four recombinant house dust mite group 1 allergens, Der p 1-N52Q, Der p 1-WT, Der f 1-N53Q, and Der f 1-WT, were secreted into the culture supernatant of transfectant cells of Pichia pastoris and converted to the mature forms with prosequences removed by dialysis against an acidic buffer.…”

Section: Methodsmentioning

confidence: 99%

The Squamous Cell Carcinoma Antigen 2 Inhibits the Cysteine Proteinase Activity of a Major Mite Allergen, Der p 1

Sakata¹,

Arima²,

Takai

et al. 2004

Journal of Biological Chemistry

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The squamous cell carcinoma antigens 1 (SCCA1) and SCCA2 belong to the ovalbumin-serpin family. Although SCCA1 and SCCA2 are closely homologous, these two molecules have distinct properties; SCCA1 inhibits cysteine proteinases such as cathepsin K, L, and S, whereas SCCA2 inhibits serine proteinases such as cathepsin G and human mast cell chymase. Although several intrinsic target proteinases for SCCA1 and SCCA2 have been found, the biological roles of SCCA1 and SCCA2 remain unknown. A mite allergen, Der p 1, is one of the most immunodominant allergens and also acts as a cysteine proteinase probably involved in the pathogenesis of allergic diseases. We have recently shown that both SCCA1 and SCCA2 are induced by two related Th2-type cytokines, IL-4 and IL-13, in bronchial epithelial cells and that SCCA expression is augmented in bronchial asthma patients. In this study, we explored the possibility that SCCA proteins target Der p 1, and it turned out that SCCA2, but not SCCA1, inhibited the catalytic activities of Der p 1. We furthermore analyzed the inhibitory mechanism of SCCA2 on Der p 1. SCCA2 contributed the suicide substrate-like mechanism without formation of a covalent complex, causing irreversible impairment of the catalytic activity of Der p 1, as SCCA1 does on papain. In addition, resistance to cleavage by Der p 1 also contributed to the inhibitory mechanism of SCCA2. These results suggest that SCCA2 acts as a crossclass serpin targeting an extrinsic cysteine proteinase derived from house dust mites and that it may have a protective role against biological reactions caused by mites.

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“…Recombinant Der p 1 and Der f 1 were prepared as previously described (29,30), with some modifications. Briefly, proforms of recombinant proteins were secreted into the culture supernatant of yeast Pichia pastoris and converted into their prosequence-removed mature forms, Der p 1-N52Q (rDer p 1), Der f 1-N53Q (rDer f 1), Der f 1-WT, and rDer f 1-C35S (11,29,30). The mature forms were then purified by anion exchange column and sizeexclusion column chromatography.…”

Section: Agsmentioning

confidence: 99%

IL-33–Mediated Innate Response and Adaptive Immune Cells Contribute to Maximum Responses of Protease Allergen–Induced Allergic Airway Inflammation

Kamijo

Takeda

Tokura

et al. 2013

The Journal of Immunology

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154

164

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How the innate and adaptive immune systems cooperate in the natural history of allergic diseases has been largely unknown. Plant-derived allergen, papain, and mite allergens, Der f 1 and Der p 1, belong to the same family of cysteine proteases. We examined the role of protease allergens in the induction of Ab production and airway inflammation after repeated intranasal administration without adjuvants and that in basophil/mast cell stimulation in vitro. Papain induced papain-specific IgE/IgG1 and lung eosinophilia. Der f 1 induced Der f 1–specific IgG1 and eosinophilia. Although papain-, Der f 1–, and Der p 1–stimulated basophils expressed allergy-inducing cytokines, including IL-4 in vitro, basophil-depleting Ab and mast cell deficiency did not suppress the papain-induced in vivo responses. Protease inhibitor–treated allergens and a catalytic site mutant did not induce the responses. These results indicate that protease activity is essential to Ab production and eosinophilia in vivo and basophil activation in vitro. IL-33–deficient mice lacked eosinophilia and had reduced papain-specific IgE/IgG1. Coadministration of OVA with papain induced OVA-specific IgE/IgG1, which was reduced in IL-33–deficient mice. We demonstrated IL-33 release, subsequent IL-33–dependent IL-5/IL-13 release, and activation of T1/ST2-expressing lineage−CD25+CD44+ innate lymphoid cells in the lung after papain inhalation, suggesting the contribution of the IL-33–type 2 innate lymphoid cell–IL-5/IL-13 axis to the papain-induced airway eosinophilia. Rag2-deficient mice, which lack adaptive immune cells, showed significant, but less severe, eosinophilia. Collectively, these results suggest cooperation of adaptive immune cells and IL-33–responsive innate cells in protease-dependent allergic airway inflammation.

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Maturation of the activities of recombinant mite allergens Der p 1 and Der f 1, and its implication in the blockade of proteolytic activity

Cited by 44 publications

References 49 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

The Squamous Cell Carcinoma Antigen 2 Inhibits the Cysteine Proteinase Activity of a Major Mite Allergen, Der p 1

IL-33–Mediated Innate Response and Adaptive Immune Cells Contribute to Maximum Responses of Protease Allergen–Induced Allergic Airway Inflammation

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