Molecular and immunological characterization of subtilisin like serine protease, a major allergen of Curvularia lunata

Tripathi, Prabhanshu; Nair, Smitha; Singh, Bhanu; Arora, Naveen

doi:10.1016/j.imbio.2010.06.009

Cited by 17 publications

(12 citation statements)

References 37 publications

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“…The biological activity of recombinant allergens was determined by basophil histamine release assay (14,25). In this study, rPer a 10 and nPer 10 induced significant histamine release by 47 to 64% and 60 to 85%, respectively, which was positively correlated.…”

Section: Discussionmentioning

confidence: 68%

“…The purified protein was fractionated on 12% polyacrylamide gel containing copolymerized 0.1% gelatin as the substrate. The gels were washed with 2.5% (vol/vol) Triton X-100 for 1 h on an orbital shaker to remove the SDS, as described previously (14). The gels were then rinsed twice with deionized water to remove the Triton X-100 and incubated overnight at 37°C in 100 mM Tris-glycine (pH 7.4), stained with CBB, and destained.…”

Section: Methodsmentioning

confidence: 99%

“…Recently a major allergen from C. lunata was cloned, expressed, and characterized as subtilisin-like serine protease (14). Recombinant DNA technology has provided the opportunity to study the specific allergenic proteins, which may be modified to reduce allergenicity for safer treatments (15,16).…”

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confidence: 99%

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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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Section: Discussionmentioning

confidence: 68%

Section: Methodsmentioning

confidence: 99%

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Characterization of Recombinant Per a 10 from Periplaneta americana

Govindaraj

Gaur

Arora

2013

Clin Vaccine Immunol

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“…Subtilisin, one of the serine proteases, have been detected from ES proteins of Acanthamoeba, and also it was known as an inducer of asthma, [22,23]. In addition, subtilisin has been detected from various organisms, and it can stimulate specific antibodies production in mice, and elicit various allergic response [24,25].…”

Section: Acanthamoeba Produced Strong Es Proteases That Could Induce mentioning

confidence: 99%

Free-Living Amoeba Acanthamoeba Triggers Allergic Inflammation of Airways

Yu¹

2015

Allergic Diseases - New Insights

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“…Evidence suggests that intrinsic biochemical activity of allergens plays a role in sensitization. Certain potent allergens from cockroaches, HDM, fungi and pollens have been shown to possess proteolytic activity [2][3][4] . Protease allergens enhance IgE responses and play a part in allergen presentation.…”

Section: Both Endogenous and Exogenous Proteases Have Been Implicatedmentioning

confidence: 99%

A cockroach derived serine protease allergen activates airway epithelial cells via PAR2 receptors

Kale

Arora

2014

Inflamm Cell Signal

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Both endogenous and exogenous proteases have been implicated in allergic airway diseases. Though recent studies have demonstrated the role of airway epithelial cells in pathophysiology of allergic diseases, the mechanisms of epithelial activation remain largely unknown. Recently published study (Kale and Arora, 2014) focused on elucidating the role of proteolytic activity of Per a 10 on airway epithelial activation and to gain an insight into possible mechanism. We found that airway epithelial activation by Per a 10 is dependent on its serine protease activity. Further we showed that this activation is PAR2 dependent and leads to Ca 2+ mobilisation. This Research Highlight discusses the findings of our recent study and active research endeavors.

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Molecular and immunological characterization of subtilisin like serine protease, a major allergen of Curvularia lunata

Cited by 17 publications

References 37 publications

Characterization of Recombinant Per a 10 from Periplaneta americana

Characterization of Recombinant Per a 10 from Periplaneta americana

Free-Living Amoeba Acanthamoeba Triggers Allergic Inflammation of Airways

A cockroach derived serine protease allergen activates airway epithelial cells via PAR2 receptors

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