cDNA cloning, sequence analysis and allergological characterization of Par j 2.0101, a new major allergen of the Parietaria judaica pollen

Duro, Giovanni; Colombo, Piergiuseppe; Costa, Maria Assunta; Izzo, V.; Porcasi, Rossana; Fiore, Renata Di; Locorotondo, Giovanni; Mirisola, Mario G.; Cocchiara, Roberta; Geraci, Domenico

doi:10.1016/s0014-5793(96)01328-2

Cited by 51 publications

(46 citation statements)

References 13 publications

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“…In particular, Parietaria judaica (Pj) pollen represents one of the main sources of allergens in the Mediterranean area [11], and Pj IgE-specific antibodies have also been detected in certain areas of southern UK, America and Australia [12]. The composition of allergenic extracts of Pj pollen has been extensively studied, and the two major allergens and isoforms have been cloned and immunologically characterised [13, 14, 15, 16, 17, 18, 19]. The Par j 1.0102 molecule is a 176-amino acid protein with a 37-amino acid NH 2 terminal signal peptide giving a 139-amino acid mature processed protein with a deduced molecular mass of 14,400 D. It is a major allergen, since it induces a response in 95% of Pj-allergic patients, and the recombinant protein produced in Escherichia coli shows a pattern of reaction similar to the native counterpart [17, 18, 20].…”

Section: Introductionmentioning

confidence: 99%

Hypoallergenic Variants of the Parietaria judaica Major Allergen Par j 1: A Member of the Non-Specific Lipid Transfer Protein Plant Family

Bonura¹,

Amoroso²,

Locorotondo³

et al. 2001

Int Arch Allergy Immunol

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Background: Par j 1 represents a major allergenic component of Parietaria judaica (Pj) pollen, since it is able to induce an immunoglobulin E (IgE) response in 95% of Pj-allergic patients. It belongs to the non-specific lipid transfer protein family, sharing with them a common three-dimensional structure. Methods: Disulphide bond variants of the recombinant Par j 1 (rPar j 1) allergen were generated by site-directed mutagenesis, and the immunological activity of rPar j 1 and its conformational mutants was compared with the use of the skin prick test (SPT). The ability to bind IgE antibodies was evaluated by Western blot, ELISA and ELISA inhibition. T cell reactivity was measured by peripheral blood mononuclear cell proliferation assay. Results: The disruption of Cys14–Cys29 and Cys30–Cys75 bridging (PjA mutant) caused the loss of the majority of specific IgE-binding activity. Additional disruption of the Cys4–Cys52 bridge (PjC mutant) and the latter Cys50–Cys91 bridge (PjD mutant) led to the abolition of IgE-binding activity. On the SPT, PjB (lacking the Cys4–Cys52 and Cys50–Cys91 bridges) was still capable of triggering a type I hypersensitive reaction in 9 out of 10 patients, and PjA in 3 out of 10 patients, while PjC and PjD did not show any SPT reactivity. All the mutants preserved their T cell reactivity. Conclusion: Recombinant hypoallergenic variants of the rPar j 1 allergen described herein may represent a useful tool for improved immunotherapy.

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Section: Introductionmentioning

confidence: 99%

Hypoallergenic Variants of the Parietaria judaica Major Allergen Par j 1: A Member of the Non-Specific Lipid Transfer Protein Plant Family

Bonura¹,

Amoroso²,

Locorotondo³

et al. 2001

Int Arch Allergy Immunol

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“…U14168), barley [34] and wheat [28]. Another homologous sequence which appears to be a member of this family is derived from a major allergen in Parietaria [29]. A common feature shared by Amb a 6 and all other members of this lipid transfer protein family which we examined (10 random members of varying homologies) is the presence of eight highly conserved cysteine residues presumably involved in four disulfide bonds.…”

Section: Discussionmentioning

confidence: 99%

“…᭧ 1998 Blackwell Science Ltd, Scandinavian Journal of Immunology, 48, [26][27][28][29][30][31][32][33][34][35][36] days, a sample of supernatant was taken from each flask and 0.5% MeOH was added to each. rAmb a 6 production was monitored by SDS-PAGE and Western blot analysis.…”

Section: Cloning and Expression Of Amb A 6 31mentioning

confidence: 99%

Cloning and Expression of Ragweed Allergen Amb a 6

Lubahn

1998

Scand J Immunol

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Hiller KM, Lubahn BC, Klapper DG. Cloning and Expression of Ragweed Allergen Amb a 6. Scand J Immunol 1998;48:26-36 We have cloned the protein coding region of an isoform of short ragweed allergen Amb a 6 (Ra6) and expressed the secreted product in Pichia pastoris at mg/l levels. 5 0 RACE was performed using sequence obtained from a partial Amb a 6 clone. This yielded a product whose deduced protein sequence has a characteristic signal sequence motif at the N-terminus followed by sequence consistent with that previously published for highly purified Amb a 6 [Roebber et al. J Immunol 1983;131:706-11]. The region encoding the secreted product was amplified by PCR and cloned into pPICZaa, an expression vector for the yeast Pichia pastoris. Yeast transformed with this vector secrete a protein which migrates near Amb a 6 in SDS-PAGE. This secreted protein reacts with polyclonal anti-Amb a 6 antisera as well as an anti-Amb a 6 monoclonal antibody, and has the N-terminal sequence of Amb a 6. By time-of-flight mass spectrometry, recombinant Amb a 6 has a molecular weight of 9884 Ϯ 0.2%. In addition to the deduced amino acid sequence of an Amb a 6 clone, the amino acid sequence of Amb a 6 protein is reported for comparison. The amino acid sequence was obtained by aligning overlapping tryptic and chymotryptic peptides from enzymatic digests of extensively reduced and alkylated Amb a 6. Sequences from at least three closely related Amb a 6 isoforms are present among these peptides. The amino acid sequence closely matches the deduced amino acid sequence of the Amb a 6 clone.

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“…judaica es una de las fuentes más importante de polen alérgeno en la zona Mediterránea (Stumvoll et al, 2003;Fotiu et al, 2011). La mayoría de trabajos existentes sobre esta especie han sido llevados a cabo para caracterizar los alérge-nos presentes en su polen (Ford et al, 1986;Costa et al, 1994;Duro et al, 1996;Stumvoll et al, 2003), además de estudiar distintos factores que condicionan la concentración del mismo en el ambiente (Fornaciari et al, 1992;Galán et al, 2000;Fotiu et al, 2011).…”

Section: Parietaria Judaica Lunclassified

Fitotoxicidad De Aceites Esenciales Y Extractos Acuosos De Plantas Mediterráneas Para El Control De Arvenses

Sancho¹,

Mercedes²

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cDNA cloning, sequence analysis and allergological characterization of Par j 2.0101, a new major allergen of the Parietaria judaica pollen

Cited by 51 publications

References 13 publications

Hypoallergenic Variants of the <i>Parietaria judaica</i> Major Allergen Par j 1: A Member of the Non-Specific Lipid Transfer Protein Plant Family

Hypoallergenic Variants of the <i>Parietaria judaica</i> Major Allergen Par j 1: A Member of the Non-Specific Lipid Transfer Protein Plant Family

Cloning and Expression of Ragweed Allergen Amb a 6

Fitotoxicidad De Aceites Esenciales Y Extractos Acuosos De Plantas Mediterráneas Para El Control De Arvenses

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