Novel Allergen Structures with Tandem Amino Acid Repeats Derived from German and American Cockroach

Pomés, Anna; Melén, Erik; Vailes, Lisa D.; Retief, Jacques D.; Arruda, L. Karla; Chapman, Martin D.

doi:10.1074/jbc.273.46.30801

Cited by 99 publications

(105 citation statements)

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“…Inserts encoding for one, two and seven duplexes were obtained by PCR. Inserts with one and two duplexes were simultaneously amplified using the Bla g 1.0101 cDNA that encodes for two duplexes as template, and an N-terminus primer encoding for the sequence GLTL NAKA (which is the beginning of one duplex) [7]. The inserts were separated in a 1% agarose gel and purified.…”

Section: A T E R I a L S A N D M E T H O D Smentioning

confidence: 99%

“…): MG.KSIPSTR. Therefore, the insert would encode for the full protein including the N-terminus [6,7]. Vectors pET 22b(+) and pET 21d(+) were digested and inserts were ligated into the NcoI and XhoI sites.…”

Section: A T E R I a L S A N D M E T H O D Smentioning

confidence: 99%

“…The novel structural feature of the group 1 allergens is that they comprise multiple tandem repeats of 100 aminoacid residues and appear to be derived from a 90 kDa precursor [6,7]. At the DNA level, the degree of homology between alternate amino-acid repeats is higher (91-95%) than between consecutive repeats (44-50%).…”

mentioning

confidence: 99%

“…In the US, this problem is particularly important in inner city areas where infestation by the German cockroach (Blattella germanica) is common [2][3][4][5]. Several German cockroach allergens have been cloned including Bla g 1, Bla g 2, Bla g 4, Bla g 5 and Bla g 6 [6][7][8][9][10]. Bla g 1 is the only German cockroach allergen that shows antigenic cross-reactivity with an American cockroach (Periplaneta americana) allergen, Per a 1 [7,[11][12][13][14].…”

mentioning

confidence: 99%

“…Several German cockroach allergens have been cloned including Bla g 1, Bla g 2, Bla g 4, Bla g 5 and Bla g 6 [6][7][8][9][10]. Bla g 1 is the only German cockroach allergen that shows antigenic cross-reactivity with an American cockroach (Periplaneta americana) allergen, Per a 1 [7,[11][12][13][14]. Bla g 1 and Per a 1 share 70% sequence identity, and the prevalence of IgE antibodies to the group I allergens in cockroach allergic patients is 30-50%.…”

mentioning

confidence: 99%

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IgE reactivity of tandem repeats derived from cockroach allergen, Bla g 1

Pomés

Vailes

Helm

et al. 2002

European Journal of Biochemistry

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Sensitization to cockroach allergens is associated with the development of asthma. Bla g 1 is a German cockroach allergen that shows allergenic cross‐reactivity with American cockroach allergen, Per a 1, and has a molecular structure composed of multiple tandem amino‐acid repeats. Two consecutive repeats are not identical but form a duplex that constitutes a basic molecular unit of Bla g 1. By molecular mass, purified natural Bla g 1 would contain approximately two duplexes. We investigated the pattern of IgE antibody binding to this repeated structure, and whether one or two duplexes are sufficient for IgE binding. Recombinant (r)Bla g 1 duplexes were expressed in Escherichia coli and in Pichia pastoris, and analyzed for monoclonal antibody and IgE antibody binding by ELISA and/or immunoblotting. Optimal rBla g 1 expression was obtained using methanol‐inducible P. pastoris (> 95% pure protein, yield ≈ 48 mg·L−1), and rBla g 1 was produced as multiple molecular forms of molecular mass 43, 32, 21 and 6 kDa, that were the result of proteolytic cleavage. There was an excellent correlation between IgE antibody binding to natural and recombinant Bla g 1 (r = 0.91, n = 29, P < 0.001), and immunoblot analysis showed that a single Bla g 1 duplex was sufficient for IgE antibody binding. The rBla g 1 is suitable for structural studies and a candidate for clinical use in diagnosis of cockroach allergy and development of new forms of immunotherapy.

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Section: A T E R I a L S A N D M E T H O D Smentioning

confidence: 99%

Section: A T E R I a L S A N D M E T H O D Smentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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IgE reactivity of tandem repeats derived from cockroach allergen, Bla g 1

Pomés

Vailes

Helm

et al. 2002

European Journal of Biochemistry

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Cockroach, Tick, Storage Mite, and Other Arthropod Allergies: Molecular Aspects

Hilger

Kuehn

Raulf

et al. 2017

Molecular Allergy Diagnostics

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A targeted approach to the identification of candidate genes determining susceptibility to Plasmodium gallinaceum in Aedes aegypti

et al. 2003

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The malaria parasite, Plasmodium, has evolved an intricate life cycle that includes stages specific to a mosquito vector and to the vertebrate host. The mosquito midgut represents the first barrier Plasmodium parasites encounter following their ingestion with a blood meal from an infected vertebrate. Elucidation of the molecular interaction between the parasite and the mosquito could help identify novel approaches to preventing parasite development and subsequent transmission to vertebrates. We have used an integrated Bulked Segregant Analysis-Differential Display (BSA-DD) approach to target genes expressed that are in the midgut and located within two genome regions involved in determining susceptibility to P. gallinaceum in the mosquito Aedes aegypti. A total of twenty-two genes were identified and characterized, including five genes with no homologues in public sequence databases. Eight of these genes were mapped genetically to intervals on chromosome 2 that contain two quantitative trait loci (QTLs) that determine susceptibility to infection by P. gallinaceum. Expression analysis revealed several expression patterns, and ten genes were specifically or preferentially expressed in the midgut of adult females. Real-time PCR quantification of expression with respect to the time of blood meal ingestion and infection status in mosquito strains permissive and refractory for malaria revealed a differential expression pattern for seven genes. These represent candidate genes that may influence the ability of the mosquito vector to support the development of Plasmodium parasites. Here we describe their isolation and discuss their putative roles in parasite-mosquito interactions and their use as potential targets in strategies designed to block transmission of malaria.

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Novel Allergen Structures with Tandem Amino Acid Repeats Derived from German and American Cockroach

Cited by 99 publications

References 28 publications

IgE reactivity of tandem repeats derived from cockroach allergen, Bla g 1

IgE reactivity of tandem repeats derived from cockroach allergen, Bla g 1

Cockroach, Tick, Storage Mite, and Other Arthropod Allergies: Molecular Aspects

A targeted approach to the identification of candidate genes determining susceptibility to Plasmodium gallinaceum in Aedes aegypti

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