Isolation and preliminary characterization of cDNA encoding American cockroach allergens

Wu, Chun-Ying; Lee, Mey-Fann; Liao, Sin-Chung

doi:10.1016/s0091-6749(95)70054-4

Cited by 29 publications

(13 citation statements)

References 24 publications

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“…Subsequently, several clones recognized by anti-Cr-PI or antiCr-PII mAb and human IgE antibodies were identified. Recombinant proteins of M r 78,000 and 72,000 from clones C12 and C20, respectively, were recognized by both anti-Cr-PI mAb and human-specific IgE antibodies (30). The cDNA sequences from C12 and C20 inserts are presented in Fig.…”

Section: Resultsmentioning

confidence: 99%

“…Sequencing of cDNA-IgE-binding and anti-Cr-PI mAb-reactive cDNA clones C12 and C20 were isolated from a gt22A cDNA library derived from the American cockroach mRNA (22,30). Phage DNAs were isolated by a polyethylene glycol precipitation method (31).…”

Section: Methodsmentioning

confidence: 99%

“…An insert was ligated into the multiple cloning sites of the pET system (Novagen, Inc., Madison, WI) and used to transform E. coli BL21(DE3) (30,(35)(36)(37). Recombinant proteins were recovered under denaturing conditions from inclusion bodies of the host cells, and purification of fusion proteins under denaturing conditions was performed using rapid affinity column chromatography with the pET His-Tag system as described by Novagen.…”

Section: /Ebi Data Bank With Accession Number(s) L40818 (C12) and L40mentioning

confidence: 99%

“…Clones C12 and C20 with 2.6-and 2.4-kilobase inserts, respectively, were recognized by both anti-Cr-PI mAb and humanspecific IgE antibodies and were selected and subcloned into expression vectors and used to transform Escherichia coli. Immunoblot analysis of clones C12 and C20 revealed fusion proteins of M r 78,000 and 72,000, respectively, recognized by both anti-Cr-PI mAb and human-specific IgE antibodies (30). In this article we report the complete nucleotide and deduced amino acid sequences of these two cDNA clones encoding the major American cockroach allergens.…”

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confidence: 97%

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Sequencing Analysis of cDNA Clones Encoding the American Cockroach Cr-PI Allergens

Lee

Liao

et al. 1996

Journal of Biological Chemistry

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A previous article described the isolation of several gt22A cDNA clones expressing the American cockroach (Periplaneta americana) Cr-PI allergens recognized by both human atopic IgE antibodies and anti-Cr-PI monoclonal antibodies (Wu, C. H., Lee, M. F., and Liao, S. C. (1995) J. Allergy Clin. Immunol. 96, 352-359). This article presents the nucleotide and deduced amino acid sequences of two cDNA clones encoding major allergens of P. americana. Clones C12 and C20 encode proteins of 685 and 631 amino acids with two potential N-glycosylation sites each. The predicted molecular weights for C12 and C20 cloned proteins are 79,300 and 75,500 with isoelectric point values of 6.26 and 6.63, which are compatible with the determined sizes (M r 78,000 and 72,000) and isoelectric point value (6.2) of the Cr-PI allergens of P. americana. A high degree of identity (69.1%), including several overlapped predicted central antigenic determinant residues, was found between two allergens. The anti-fusion protein antibody-based enzyme-linked immunosorbent assay was able to detect crude American cockroach extract, Cr-PI, recombinant proteins, and commercial cockroach extracts, which provides further evidence that two allergens share common antigen determinants. Recombinant allergens of clones C12 and C20 both showed 47.4% skin reactivities on 19 cockroach-sensitive asthmatic patients. Unexpectedly, although no sequence similarity was found to other known allergens, two aromatic amino acid-rich allergens were found to have a striking sequence identity to insect storage proteins (20.1-33.9%), insect juvenile hormone-suppressible proteins (30.9 -36.4%), and arthropod hemocyanins (29.7-34.6%). Results suggested that two prominent allergens of P. americana are ancestrally related to these insect hemolymph proteins and represent a new group of proteins in the hemocyanin superfamily. These data will now facilitate epitope-mapping studies, and the recombinant allergens may be valuable for diagnostic and therapeutic purposes.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: /Ebi Data Bank With Accession Number(s) L40818 (C12) and L40mentioning

confidence: 99%

mentioning

confidence: 97%

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Sequencing Analysis of cDNA Clones Encoding the American Cockroach Cr-PI Allergens

Lee

Liao

et al. 1996

Journal of Biological Chemistry

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“…Current evidence suggests that B. germanica produces at least five allergens that elicit IgE responses in 30 -70% of CR-allergic patients (11-13, 33, 34). A high molecular mass allergen (72 kDa) has recently been cloned from P. americana, designated Per a 3; however, the homologous allergen does not appear to occur in B. germanica (35). Cloning of high molecular mass B. germanica allergens has also been reported (36).…”

Section: Discussionmentioning

confidence: 99%

Cloning of Cockroach Allergen, Bla g 4, Identifies Ligand Binding Proteins (or Calycins) as a Cause of IgE Antibody Responses

Arruda

Vailes

Hayden

et al. 1995

Journal of Biological Chemistry

130

118

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An allergen cloned from a Blattella germanica (German cockroach) cDNA library, encoded a 182-amino acid protein of 20,904 Da. This protein, designated B. germanica allergen 4 (Bla g 4), was expressed as a glutathione S-transferase fusion protein in Escherichia coli and purified by affinity chromatography and high-performance liquid chromatography. The prevalence of serum IgE antibody to recombinant Bla g 4 in 73 cockroach allergic patients with asthma ranged from 40% (antigen binding radioimmunoassay) to 60% (plaque immunoassay). Cockroach allergic patients gave positive intradermal skin tests to recombinant Bla g 4 at concentrations of 10 ؊3 -10 ؊5 g/ml, whereas non-allergic controls, or cockroach allergic patients with no detectable serum IgE antibody to Bla g 4, gave negative skin tests to 1 g/ml. Polymerase chain reaction and Southern analysis identified a 523-base pair DNA encoding Bla g 4 in both B. germanica and Periplaneta americana (American cockroach). However, Northern analysis showed that mRNA encoding Bla g 4 was transcribed in B. germanica but not in P. americana, suggesting that allergen expression was species specific. Sequence similarity searches showed that Bla g 4 was a ligand binding protein or calycin and unexpectedly revealed that this family contained several important allergens: ␤-lactoglobulin, from cow milk, and rat and mouse urinary proteins. Although the overall sequence homology between these proteins was low (ϳ20%), macromolecular modeling techniques were used to generate two models of the tertiary structure of Bla g 4, based on comparisons with the x-ray crystal coordinates of bilin binding protein and rodent urinary proteins. The results show that members of the calycin protein family can cause IgE antibody responses by inhalation or ingestion and are associated with asthma and food hypersensitivity.

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Immunology

2008

Pediatric Allergy, Asthma and Immunology

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Isolation and preliminary characterization of cDNA encoding American cockroach allergens

Cited by 29 publications

References 24 publications

Sequencing Analysis of cDNA Clones Encoding the American Cockroach Cr-PI Allergens

Sequencing Analysis of cDNA Clones Encoding the American Cockroach Cr-PI Allergens

Cloning of Cockroach Allergen, Bla g 4, Identifies Ligand Binding Proteins (or Calycins) as a Cause of IgE Antibody Responses

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