Molecular and immunological characterization of Pen ch 18, the vacuolar serine protease major allergen of Penicillium chrysogenum

Shen, Horng Der; Chou, Hong; Tam, Ming F.; Chang, Ching-Yun; Lai, Hsin-Hua; Wang, S.-R.

doi:10.1034/j.1398-9995.2003.00107.x

Cited by 54 publications

(50 citation statements)

References 29 publications

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“…Besides N-terminal cleavage, the mature vacuolar serine proteases may also undergo C-terminal processing [15, 20]. Similar to that reported for the vacuolar serine protease from C. albicans [17], the deduced cDNA sequence of Rho m 2 does not have a C-terminal extension.…”

Section: Discussionmentioning

confidence: 56%

“…IgE cross-reactivity between the group 18 (Pen ch 18) and the group 13 (Pen ch 13) major allergens from P. chrysogenum has been reported [20]. In addition, we have also demonstrated IgE cross-reactivity among serine protease allergens from P. oxalicum, P. chrysogenum and P. citrinum [22], as well as from A. fumigatus , P. chrysogenum and P. oxalicum [23].…”

Section: Discussionmentioning

confidence: 68%

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A Vacuolar Serine Protease (Rho m 2) Is a Major Allergen of Rhodotorula mucilaginosa and Belongs to a Class of Highly Conserved Pan-Fungal Allergens

Chou¹,

Tam²,

Lee³

et al. 2005

Int Arch Allergy Immunol

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Background:Rhodotorula mucilaginosa is one of the most frequently encountered species of yeasts in our environment. We reported here a major allergen of R. mucilaginosa. Methods: A major R. mucilaginosa allergen (Rho m 2) was characterized by two-dimensional (2D) immunoblotting, protein sequencing, cDNA cloning and IgE cross-reactivity with fungal serine proteases. Results: Fourty-four sera (28%) from 157 bronchial asthmatic patients showed IgE-immunoblot reactivity against R. mucilaginosa extract. Among these 44 sera, 25 (57%) demonstrated IgE binding against a 31-kDa protein of R. mucilaginosa. Protein sequencing results suggest that it is a vacuolar serine protease. The corresponding cDNA clone encoding a mature protein of 312 residues was isolated. It shares 67–68% sequence identity with vacuolar serine protease allergens from three different Penicillium species (Pen ch 18, Pen o 18 and Pen c 18) and designated as Rho m 2 by the Allergen Nomenclature Committee. The native and recombinant Rho m 2 react with IgE antibodies and monoclonal antibody (MoAb) FUM20 against fungal serine proteases. IgE cross-reactivity between nRho m 2 and nPen ch 18 was observed. It was also detectable between rRho m 2 and rPen o 18. Conclusion: Our results suggest that R. mucilaginosa may also be a significant causative agent of human respiratory allergic disorders. We identified a vacuolar serine protease as a major allergen of R. mucilaginosa (Rho m 2) and a pan allergen of prevalent airborne fungal species. We detected IgE cross-reactivity among these highly conserved serine protease pan-fungal allergens.

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Section: Discussionmentioning

confidence: 56%

Section: Discussionmentioning

confidence: 68%

A Vacuolar Serine Protease (Rho m 2) Is a Major Allergen of Rhodotorula mucilaginosa and Belongs to a Class of Highly Conserved Pan-Fungal Allergens

Chou¹,

Tam²,

Lee³

et al. 2005

Int Arch Allergy Immunol

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“…Furthermore, IgE binding of rPer a 10 was completely inhibited when pooled patients' sera were preabsorbed with 500 g of self-protein in immunoblot inhibition, and there was no inhibition in C. lunata serine protease, which showed clear bands. Serine proteases are reported from many sources, and most of them are cross-reactive allergens (22)(23)(24).…”

Section: Discussionmentioning

confidence: 99%

Characterization of Recombinant Per a 10 from Periplaneta americana

Govindaraj

Gaur

Arora

2013

Clin Vaccine Immunol

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bCockroach allergen is a major risk factor for IgE-mediated allergic response and asthma in sensitized individuals. Serine proteases have been identified from various sources and characterized as major allergens. The present study was aimed to express and characterize recombinant allergen Per a 10 (rPer a 10) from Periplaneta americana. rPer a 10 was expressed in Escherichia coli and purified in soluble form, yielding 0.75 mg/liter of culture. Homology of the Per a 10 protein sequence exhibited 27 to 38% similarity to the mite serine protease and 41 to 52% similarity to other insect trypsins. The purified rPer a 10 protein resolved at 28 kDa on SDS-PAGE and was recognized by cockroach-hypersensitive patients' sera by immunoblotting and enzymelinked immunosorbent assay (ELISA). In competitive ELISA, rPer a 10 required 96 ng of purified protein for 50% inhibition of IgE binding, whereas 34 ng of native protein (nPer a 10) was required for the same inhibition. rPer a 10 and nPer a 10 induced basophil histamine release in the range of 47 to 64% and 60 to 85%, respectively, when sensitized with cockroach-hypersensitive patients' sera. In conclusion, Per a 10 was subcloned, and the protein was purified to homogeneity. rPer a 10 showed reduced IgE binding and histamine release and showed no proteolytic activity. These data suggest that rPer a 10 has potential for immunotherapy.

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“…We have previously reported that FUM20 can bind a synthetic peptide with sequence encompassing the N-terminal 23 residues of the mature Pen ch 18 [11]. mAb FUM20 also recognizes a class of pan-fungal allergens.…”

Section: Introductionmentioning

confidence: 99%

Lys, Pro and Trp Are Critical Core Amino Acid Residues Recognized by FUM20, a Monoclonal Antibody against Serine Protease Pan-Fungal Allergens

Lee

Tam²,

Chou³

et al. 2007

Int Arch Allergy Immunol

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Background: Alkaline/vacuolar serine proteases comprise a major group of pan-fungal allergens from several prevalent airborne fungal species. It is of importance to characterize antigenic determinant(s) recognized by monoclonal antibodies against these major allergens. Methods: The antigenic determinant of fungal serine proteases recognized by a monoclonal antibody, FUM20, was analyzed by dot immunoassay of synthetic peptides immobilized on cellulose membrane. Results obtained were confirmed by wild-type recombinant protease and its mutants. The epitopes were mapped to the structure of serine proteases by molecular modeling. Results: A linear epitope encompassing 9 amino acids from Pen ch 18 (⁶EKNAPWGLA¹⁴) binds FUM20. The corresponding peptide (⁵AKGAPWGLA¹³) from Rho m 2 also binds FUM20. Substitution of K6, P9 or W10 with alanine in this peptide resulted in drastic loss of FUM20 binding. Rho m 2 mutants with single K6A, P9A, P9G, W10A or W10F substitute showed negative immunoblot reactivity against FUM20. However, the Rho m 2 K6R mutant can bind FUM20. Three-dimensional structural models of the FUM20 antigenic determinants on serine proteases were constructed. The lysine residue critical for FUM20 interaction is on the surface of the proteases and solvent accessible. The critical core residue proline is located at the beginning of an α-helix. Conclusions: The lysine, proline and tryptophan residues located on the N-terminal region of fungal serine proteases are critical core amino acid residues recognized by FUM20, a monoclonal antibody against serine protease pan-fungal allergens. These findings advance our understanding of the antigenic structures responsible for the antigenicity of serine protease allergens.

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Molecular and immunological characterization of Pen ch 18, the vacuolar serine protease major allergen of Penicillium chrysogenum

Cited by 54 publications

References 29 publications

A Vacuolar Serine Protease (Rho m 2) Is a Major Allergen of <i>Rhodotorula mucilaginosa </i>and Belongs to a Class of Highly Conserved Pan-Fungal Allergens

A Vacuolar Serine Protease (Rho m 2) Is a Major Allergen of <i>Rhodotorula mucilaginosa </i>and Belongs to a Class of Highly Conserved Pan-Fungal Allergens

Characterization of Recombinant Per a 10 from Periplaneta americana

Lys, Pro and Trp Are Critical Core Amino Acid Residues Recognized by FUM20, a Monoclonal Antibody against Serine Protease Pan-Fungal Allergens

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